+End microtubule tracking proteins (CLASPs, EB1, CLIP170, ect.)\n[[Akhmanov2005|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VRW-4F1SVTK-1&_user=4430&_coverDate=02%2F01%2F2005&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=5fe4331fbe8c1b3edd1e9135021b4a49]], [[Vaughan2004|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TCX-4D16V3G-2&_user=4430&_coverDate=09%2F01%2F2004&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=0b1792dca475d8514277540be14206f7]]\n\nDynactin and EB1 work together to regulate MT polymerization [[Ligon2003|http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=12686597]]
500 mM Tris.HCl, pH 7.4 and 1500 mM NaCl:\n\n78.8g Tris/Hcl (MW 157.6)\n87.6g NaCl (MW 58.4)\nFill to 1L with ddH20
39.4g Tris/Hcl (MW 157.6)\nFill to 250mL with ddH20
2 Photon
73.05g NaCl (MW 58.44)\nFill to 250mL with ddH2O
Dynein drives fast retrograde transport. Kinesin drives fast and ''slow'' anterograde transport - slow moves same rate as fast but pauses a lot. [[Roy2007|http://www.jneurosci.org/cgi/content/full/27/12/3131]], [[Shah2002|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VRW-44W09T8-9&_user=4430&_coverDate=02%2F01%2F2002&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=adb9688c36bfe650576c834677b9c054]]\n\nMicrotubule Transport (role of Dynein): "The occurrence of anterograde MT and retrograde NF movements was significantly diminished in neurons that had been depleted of dynein heavy chain, whereas the occurrence of retrograde MT and anterograde NF movements was unaffected. These results support a cargo model for NF transport and a sliding filament model for MT transport." [[He2005|http://www.jcb.org/cgi/content/full/168/5/697]], [[Meyers2006|http://nro.sagepub.com/cgi/reprint/12/2/107]], [[Baas2006|http://www.blackwell-synergy.com/doi/full/10.1111/j.1600-0854.2006.00392.x]]\n\nNumber of kinesins/particle affects pausing but not rate (b/c processive). Number of dyneins/particle can drastically affect rate (b/c not processive) \n[[Brown2003|http://www.jcb.org/cgi/content/full/160/6/817]], [[Duncan2006|http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=17009871]], [[Stokin2006|http://arjournals.annualreviews.org/doi/full/10.1146/annurev.biochem.75.103004.142637]]
[[Marshall2004|http://arjournals.annualreviews.org/doi/full/10.1146/annurev.cellbio.20.012103.094437]]
[[Marshall2004|http://arjournals.annualreviews.org/doi/full/10.1146/annurev.cellbio.20.012103.094437]]
Nuclear Pore\nMembrane Gating - tight junctions [[Shin2006|http://arjournals.annualreviews.org/doi/full/10.1146/annurev.cellbio.22.010305.104219]]
What oscillators operate on the ~4Hz scale? in vitro Kinase Circuits?
[[Biorad Protocol|http://www.fhcrc.org/science/labs/hahn/methods/biochem_meth/biorad_assay.pdf]]\n\n__Standards:__ \n\n1) Dilute 1 mg/ml BSA stock 1:10 to get 0.1 mg/ml BSA\n\n''0 µg/ml:'' 0 µl + 800 µl H20\n''2.5 µg/ml:'' 25 µl + 780 µl H20\n''5 µg/ml:'' 50 µl + 760 µl H20\n''7.5 µg/ml:'' 75 µl + 740 µl H20\n''10 µg/ml:'' 100 µl + 700 µl H20\n\n2) Add 200 µl concentrated Biorad reagent and incubate at room temperature for 5 minutes.\n\n3) Assay absorbance on spec at 595 nm (Bradford setting on spec). Make standard curve to fit samples. On spec, can use "parameter -> std" to measure standards and calculate concentartions of samples.\n\n[img[alternate text|\nhttp://docs.google.com/File?id=dd397pj8_435r7jtrhm]]\n\n__Samples:__\n\n1) Add 1, 2, or 4 µl of concentrated unknown (1:1000, 1:500, 1:250 dilution), and bring volume up to 800 µl with H20.\n\n2) Add 200 µl concentrated Biorad reagent and incubate at room temperature for 5 minutes.\n\n3) Assay absorbance at 595 nm.\n
+end tracking proteins that can also bind along microtubules, stabilizing the filiments.\n\n20 mM [[lithium chloride]] induces ectopic CLASP2 MT lattice binding in the cell body of PtK1 cells (CLASP2 normally only coats MTs in the lamella). Overexpression of GSK3b prevents CLASP2 lattice binding in the lamella. Constituatively active Rac1 induces ectopic CLASP2 MT binding (like lithium), wheras dominant negative Rac1 prevents CLASP2 binding (like GSK3b overexpression)\n[[Wittmann2005|http://www.jcb.org/cgi/content/full/169/6/929]], [[Torsten Wittmann]] and [[Clare Waterman-Storer]]\nInterestingly, CLASPs seem to be important in stabilizing asymmetric MTs originating at the trans-Golgi and going to leading edge. A membrane-bound protein, GCC185, recruits CLASPs to the TGN, where the CLASPs stabilize pre-existing noncentrosomal MT seeds and promote the elongation of MTs from the TGN. CLASP2 preferentially associates with Golgi MTs and CLASP RNAi prevents Golgi MT sormation after [[nocodazole]] treatment.\n[[Efimov2007|http://www.developmentalcell.com/content/article/fulltext?uid=PIIS1534580707001499]]\n[[Vaughan2002|http://www.jcb.org/cgi/content/full/158/2/305]]
10mL [[1M Tris/Hcl]] pH7.5\n14mL [[5M NaCl]]\n2mL .5M EDTA\n1g CTAB\n1mL Beta Mercapto Ethanol (14M)\n\nFill to 100mL with ddH2O
Conditionally expressed Centrin gene with the NIT1 promoter, which is repressed by NH4+ [[Schoppmeier2005|http://www3.interscience.wiley.com/cgi-bin/fulltext/110520723/PDFSTART]]
[img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_14hds6nmg3]]\n[[Naya2001|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T36-43HK96X-7&_user=4430&_coverDate=07%2F20%2F2001&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=a680b469edf0d51a1f84798b583b29cd#tbl1]]\n\n[[Codon Usage Table|http://www.kazusa.or.jp/codon/cgi-bin/showcodon.cgi?species=Chlamydomonas+reinhardtii+%5Bgbpln%5D]]
During this process, avoid contamination so plates wont grow mold:\n\n- Make gametes by picking strains into 2mL M-N media, grow overnight in light\n- Mix 2mL cultures in one tube, let sit on bench for 2 hours\n- Aliquot 500uL/plate on 4% TAP plates, shake gently to spread\n- Let dry in light (several hours or overnight)\n- Invert plates and wrap in foil (shielded from light), store for at least 5 days\n- Tetrad Dissection!
[[ts flagella loss]]\n[[short flagella]]\n[[stubby flagella / bald]]\n[[long flagella]]\n[[unequal length flagella]]\n[[paralyzed flagella]]\n[[flagellar suppressor mutations]]\n\n[[GFP tagged]]\n[[linkage groups]]
Cell wall is removed by autolysin or incubation in M-N media for ~3hrs\nVector is linearized and introduced into cells by electroporation or glass bead method, [[Shimogawara1998|http://www.genetics.org/cgi/content/full/148/4/1821]]\nTransformants are selected for with Paromycin.\n\n(Vectors include the GC rich [[aphVIII]] gene from Streptomyces rimosus, which confers resistance to Paromycin, Kanamycin, and Neomycin) [[Sizova2001|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T39-445G6X8-N&_user=4430&_coverDate=10%2F17%2F2001&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=36ae4af5a2d79b152ecc4765c8516ae1]]
Background: #fff\nForeground: #000\nPrimaryPale: #8cf\nPrimaryLight: #66ff00\nPrimaryMid: #007700\nPrimaryDark: #003300\nSecondaryPale: #bbff77\nSecondaryLight: #66cc00\nSecondaryMid: #339900\nSecondaryDark: #336600\nTertiaryPale: #cceeaa\nTertiaryLight: #66cc00\nTertiaryMid: #669900\nTertiaryDark: #666\nError: #f88\n
13kd protein enriched in basal body\n[[Schoppmeier2005|http://www3.interscience.wiley.com/cgi-bin/fulltext/110520723/PDFSTART]]
[[Hello]]
Planarian Regeneration [[SanchesAlvarado2004|http://arjournals.annualreviews.org/doi/full/10.1146/annurev.cellbio.20.010403.095114]]
Diffusion in a tube vs. Diffusion in a Membrane
A mutant of a GFP-like fluorescent protein that was cloned from the coral Pectiniidae. Intense irradiation at 488 nm changes Dronpa to a dim protonated form, but even weak irradiation at 405 nm restores it to the bright deprotonated form. Reversible photoswitching can be repeated >100 times. Fast response time on the order of milliseconds.\n[[Habuchi2005|http://www.pnas.org/cgi/content/full/102/27/9511]]
[[colchicine]]\n[[cyclohexamide]]\n[[dibucaine]]\n[[IBMX]]\n[[lithium chloride]]\n[[sodium pyrophosphate]]\n[[taxol]]\n\nDynein Inhibitors: [[purealin]], [[EHNA]]\nDynactin Inhibitors: [[dynamitin]]\nKinesin Inhibitors: [[adociasulfate-2]], [[monastrol]]\nGeneral Inhibitors: [[vanadate]]
erythro-9-[3-2-(hydrosynonyl)]adenine\nInhibits cytoplasmic and axonemal dynein ATPase activity.\n[[Penningroth1982|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WBK-4FFH1B9-86&_user=4430&_coverDate=01%2F15%2F1982&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=4d646f54ad4eb08f1faec2f34ab73a6a]]
Wrote [[The Chlamydomonas Source Book|http://www.amazon.com/Chlamydomonas-Sourcebook-Comprehensive-Biology-Laboratory/dp/012326880X]] (1989)
From the stony coral Lobophyllia hemprichii. Strong green fluorescence (516 nm) that changes to red (581 nm) upon near-UV irradiation at ~390 nm, because of a photo-induced modification involving a break in the peptide backbone next to the chromophore. Wild-type of EosFP is tetrameric, but single point mutations V123T and T158H created monomeric species that is funtional in mamalian cells. mEosFP loses fluorescence over 30C, while the dimer and tetramer forms are more stable (but worry about aggreagation).\n[[Wiedenmann2004|http://www.pnas.org/cgi/content/full/101/45/15905?ijkey=56b97321a39b715257a3f276b6b6f37a69be7c86]], [[Ivanchenko2007|http://www.biophysj.org/cgi/content/full/92/12/4451#BIB10]]\n[img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_12cjnhsjfd][http://www.pnas.org/cgi/content/full/101/45/15905/FIG2]]\n
__F__luorescence __I__maging __O__ne-__N__anometer __A__ccuracy\n\n"FIONA. A single fluorescent molecule forms a diffraction-limited image of width ~ {lambda}/2 N.A., or ~ 250 nm for visible light, where N.A. is the numerical aperture of the collection lens. The center of the image, which, under appropriate conditions, corresponds to the position of the dye, can be located to arbitrarily high precision by collecting a sufficient number of photons. Our method for determining the center relies on curve-fitting the image [also called the point-spread function (PSF)] to a Gaussian function. The fundamental goal is to determine the center or mean value of the distribution, µ = (x0, y0), and its uncertainty, the standard error of the mean, {sigma}µ. By measuring the position before and after a step, the step size can be determined." [[Yildiz2003|http://www.sciencemag.org/cgi/content/full/300/5628/2061?ijkey=5681642aa8c20537e28cc0a49f415d903912fb50]], [[Paul Selvin]]
__F__luorescence __p__hoto__a__ctivation __l__ocalization __m__icroscopy\n\n"Method for fluorescence imaging that can obtain spatial distributions of large numbers of fluorescent molecules on length ''scales shorter than the classical diffraction limit''. FPALM analyzes thousands of single fluorophores per acquisition, localizing small numbers of them at a time, at low excitation intensity. To control the number of visible fluorophores in the field of view and ensure that optically active molecules are separated by much more than the width of the point spread function, photoactivatable fluorescent molecules are used, in this case the ''photoactivatable green fluorescent protein'' ([[PA-GFP]]). For these photoactivatable molecules, the activation rate is controlled by the activation illumination intensity; nonfluorescent inactive molecules are activated by a high-frequency (405-nm) laser and are then fluorescent when excited at a lower frequency. The fluorescence is imaged by a CCD camera, and then the molecules are either ''reversibly inactivated or irreversibly photobleached'' to remove them from the field of view. The rate of photobleaching is controlled by the intensity of the laser used to excite the fluorescence, in this case an Ar+ ion laser. Because only a small number of molecules are visible at a given time, their positions can be determined precisely; with only ~100 detected photons per molecule, the ''localization precision can be as much as 10-fold better than the resolution'', depending on background levels"\n[img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_24czznwvfh][http://www.biophysj.org/cgi/content/full/91/11/4258/FIG1]]\n[[Hess2006|http://www.biophysj.org/cgi/content/full/91/11/4258]]
__PROTOCOL WRITTEN FOR TAP. M1 DIRECTIONS IN { # }__\n\n1) Grow up 100-300mL culture in TAP or M1 (bubbling) [~130mL TAP is good]\n\n2) Take ''3mL'' sample from culture to be __"whole cell extract"__. Spin down ''~1000xG, 2min @RT [GH 3.8 rotor: 2090 rpm]'', aspirate, resuspend in ''200uL'' {150uL} ~HMEDK and flash freeze in liquid nitrogen (before freeze, separate some sample {30uL} to quantify protein levels by bradford).\n \n3) Spin down all cells (whole culture) ~1000xG, 5min @RT [GH 3.8 rotor: 2090 rpm]\n\n4) Wash w/ TAP {M1} (~40mL) 2 times ~1000xG, 5min @RT\n''***this step removes cell fragments which could contaminate the flagellar fraction***''\n\n5) Resuspend w/ TAP {M1} to ''30mL''\n\n6) ''ON ICE'', [[pH Shock]] cells:\n\n - add ''1.5mL'' {600uL} of .5M acetic acid to drop to pH4\n - let sit on ice 1 min\n - add ''1.65mL'' {700uL} of .5M KOH to return to pH7.2\n\n7) Add protease inhibitors: ''300uL'' sigma protease inhibitor\n ''30uL'' 100mM EGTA\n\n8) Spin cell bodies down: ''~1000xG, 5min __@4C__ [SS 5.3 rotor: 1000 rcf]''\n\n9) Remove all supernatant and put in fresh 50mL tube\n\n10) Resuspend cell bodies in ''5mL'' {2.5mL} ~HMEDK and flash freeze some (~200uL) for __"cell bodies"__ sample (before freeze, seperate some sample to quantify).\n\n11) UNDERLAY supernatant with ''8mL'' cold 25% sucrose in TAP {M1}. \n''***use rubber plunger to suck ~11mL sucrose into 10mL pipette. Position pipette at bottom of tube and SLOWLY put in sucrose (supernatant should float on top)***''\n\n12) Spin cell bodies out: ''2500xG, 10min __@4C__ with NO BRAKE [SS 5.3 rotor: 2500 rcf]''\n\n13) Take supernatant and interface and transfer to a fresh 50mL tube\n\n14) REPEAT STEPS 11-13 \n\n15) Put Supernatant in Oak Ridge tubes and spin: ''10,000xG, 20min __@4C__ [HB 4 rotor: 10,000 rcf]''\n\n16) Remove supernatant and resuspend __"flagella"__ sample in minimal volume ~HMDEK: ''200uL'' {150uL}\n\n17) Add protease inhibitors and flash freeze (seperate out some sample for quantifying protein before freezing)\n\n18) Store all samples (whole cells, cell bodies, flagella) at ''-80C''\n\n
[[cGFP]]\n[[photoactivatable / photoconvertable FPs]]\n
[[KAP-GFP]]\n[[IFT27-GFP]]\n[[SFA-GFP]]\n[[Centrin-GFP]]\n[[DIP13-GFP]]\n\nLechtreck has failed to make constructs for alpha1-tubulin, delta-tubulin, IFT20, IFT52, IFT88, L2LIC, LC11, Rib72,\nand PF16. Can conditionally express genes with the NIT1 promoter, which is repressed by NH4+ [[Schoppmeier2005|http://www3.interscience.wiley.com/cgi-bin/fulltext/110520723/PDFSTART]]\nOverexpress with Rubisco promoters (expression varies widely with insertion site) or express wt levels w/ endogenous promoter.\n\n__''Tagged IFT proteins in other systems:''__\n__C. elegans__\nHeterotrimeric kin2: [[KAP1-GFP]]\nHomodimeric kin2: [[OSM3-GFP]]\nOSM3 kinesin cofactor: [[DYF1-GFP]]\nIFT complex linkers (linkA+B): [[BBS7-GFP]], [[BBS8-GFP]]\nIFTA: [[CHE11-GFP]] ([[IFT140]]), unlabeled- DAF10 ([[IFT122]])\nIFTB: [[CHE2-GFP]], [[OSM1-GFP]] ([[IFT172]]), [[OSM5-GFP]] ([[IFT88]]), [[OSM6-GFP]] ([[IFT52]]), [[DYF6-GFP]], unlabeled- CHE13 ([[IFT46]])\n[[Ou2005|http://www.nature.com/nature/journal/v436/n7050/full/nature03818.html]], [[Pan2006|http://www.jcb.org/cgi/content/full/174/7/1035]], [[Signor1999|http://www.jcb.org/cgi/content/full/147/3/519]], [[Bell2006|http://www.genetics.org/cgi/content/full/173/3/1275?ijkey=3121bde88730be5f427ce37d1e8a8566492d9694]]\nKymographs with Dyf-1 and IFT proteins undergoing antero and retro, but kinesin-II only going antero. However, Signor paper says that they see kinesin-II being transported retrograde. CHE3 is cytoplasmic dynein 1b.\n[[Jonathan Scholey]]\nTRPV transmembrane vanillinoid channels: [[OSM9-GFP]], [[OCR2-GFP]]\n[[QinRosenbaum2005|http://www.wisc.edu/molpharm/faculty/barr/Pub16169494.pdf]]\n\n__Dictyostemium__\nCytoplasmic Dynein: [[IC-GFP]] [[Ma2002|http://jcs.biologists.org/cgi/content/full/115/7/1453]]\n\n__S. pombe and S. cerevisiae__\nCytoplasmic Dynein: [[DHC-GFP]] [[Yamamoto199|http://www.jcb.org/cgi/content/full/jcb;145/6/1233]], [[Sha1998|http://www.molbiolcell.org/cgi/reprint/9/7/1627.pdf]]\n\n__Mammalian__\nIFTB: [[IFT20]]-GFP [[Follit2006|http://www.molbiolcell.org/cgi/reprint/17/9/3781.pdf]]
Grow 50mL of cells in [[TAP]] (grow cart shaker)\n\n__''Day 1:''__\n1) Spin down 35mL of cells @ 3000RPM, 5 min\n2) Pour off media, leaving chlamy pellet\n3) Freeze at -80C\n4) Lyphilize overnight\n\n__''Day 2:''__\n1) Vortex with glass beads (small size)\n2) Add 10mL [[CTAB Buffer]] and vortex breifly\n3) Incubate in 65C waterbath overnight\n\n__''Day 3:''__\n1) Add 10mL chloroform (and mix)\n2) Spin 5 min @ 3000RPM\n3) Transfer aqueous (top) layer to a new tube\n4) Add 10mL isopropanol (and mix)\n5) Spin 10 min @ 3000RPM\n6) Pour off supernatant\n7) Wash pellet with 10mL 70% ethanol. \n8) Spin for another 10 min @ 3000RPM\n9) Suck off supernatant and air dry on bench\n10) Resuspend pellet in 200uL TE (transfer to epi tube)\n11) Add 1uL RNAse A (10 mg/mL)\n12) Incubate 1hr @ 37C\n13) Add 50uL ProK (.5mg) (50uLof 10mg/mL = .5mg)\n14) Incubate in 55C waterbath overnight\n\n__''Day 4:''__\n1) Add 250uL phenol : chloroform (phenol:chlorophorm:isoamyl alcohol 25:24:1, 10mM Tris pH8)\n2) Spin 10 min @ 14,000RPM (cold room max speed)\n3) Transfer aqueous (top) layer to new epi tube\n4) Add 20uL [[3M sodium acetate]] (NaOAc)\n5) Add 900uL 100% Ethanol ''***PRECIPITATES DNA!***''\n6) Put in -80C for 1hr\n7) Spin 10 min @ 16,000RPM (max speed in ''cold room'' = 14,000RPM)\n8) Suck off supernatant\n9) Wash pellet with 70% ethanol (900uL)\n10) Spin 10 min @ max speed\n11) Suck off supernatant and air dry on bench\n12) Resuspend in 100uL 10% TE (or ddH2O) (or EB)\n
Type the text for 'Greg Pazour'
I am currently using this space to prepare for my quals, but hopefully it will evolve into a good resource.
isobutyl methylxanthine - a phosphodiesterase inhibitor.\n\nInduces flagellar shortening
__''DAY 1''__\n\n''1. set-up:''\na. thaw out enough blocking solution for the fIrst day. 100% block = 5% BSA plus 1 % cold water fish gelatin, in IX PBS\nb. set up two coplin jars of methanol in the -20. mark the front of each jar with a sharpie so you will know which way the coverslips are facing! \n\n''2. Prepare poly-lysine coated coverslips''\na. put 200 uL of Sigma poly lysine solution onto each coverslip (22x22), let sit10 min.\nb. dip the coverslip into a beaker of water and then let them dry on a kimwipe, poly-lysine coated face up. \n\n''3. Fix cells''\na. [optional] fix cells by adding 1:4 iodine to culture for 2 min. this is especially important for preserving flagellar proteins.\nb. put 100 uL of cells on the coverslip. the denser the culture, obviously, the more densely the cells with end up covering the coverslip.\nc. let the cells adhere for 5-10 min. if your want to get nice images of the flagella, let them adhere for less time (2 min) because the flagella start to curl up.\nd. lift up the coverslip and tap the edge against a kimwipe in order to let the excess cells and media run off.\ne. put the coverslip into a coplin jar with -20C methanol. repeat c,d for each coverslip.\n ''***whenever you put coverslips in jars, make sure you keep track of which way the coverslips are facing, i.e. where the cells are.''\nf. let the coverslips sit in the jar with methanol in the -20 freezer for 5 min.\ng. transfer the coverslips into a second coplin jar of -20C 50:50 methanol:PBS. let them sit in the second jar for 5 min. \n (these two methanol steps not only fix the cells, they also permeabilize the cell wall and extract out all the autofluorescent chlorophyll).\ni. transfer the coverslip to a coplin jar with PBS for 5 min, then pour off the PBS and add some fresh PBS, let them sit in this for 10 min. \n\n''4. Block''\na. dump out the PBS and add 100% block. let them sit in block for 30 min.\nb. dump out the block and add 10% normal goat serum made up in 100% block. tUN Uust to be clear about goat serum - you make up 100% goat serum by adding 10mL of distilled water to the bottle of serum, then you dilute this serum 1: 10 in 100% block (so it ends up being 80% block final concentration)\nc. let them sit in the block + goat serum for 30 min. \n\n''5. Stain with primary antibody''\na. make up your antibody dilutions to have a final concentration of 20% block (diluted in PBS). when diluting antibodies, calculate 100 uL per coverslip to be stained. for monoclonal antibodies, a dilution of 1 :20 usually works well. for polyclonal antibodies, 1 usually try 1 :200\nb. set up a humid chamber (I like flat tupperware boxes with a few wet kimwipes sitting inside. then place a piece of parafilm on the bottom of the box. \nc. for each coverslip, put a drop of 100 uL of antibody onto the parafilm, then invert the coverslip onto the drop.\nd. close the box and let the cells stain overnight. \ne. if you are in a hurry, an hour is probably good enough. \n\n__''DAY 2''__\n\n''6. Stain with secondary antibody''\na. wash the coverslips in 6 changes of PBS in a coplin jar. 5 min per wash (dump out the used buffer each time).\nb. make up the secondary antibody in 20% block, 1 usually dilute the secondaries 1 :500\nc. stain the covers lips by inverting them over a 100uL drop of secondary antibody, using the same procedure as with the primary except you don't need a humid chamber but you should block the room light with a box over the samples. Stain for 1 hour. During staining and subsequent washes, keep the samples in the dark so the fluorescence doesn't bleach.\nd. wash the coverslips 6 times in PBS 5 min each. \n\n''7. Stain with DAPI (to see nucleus/chromatin)''\na. stain the cells with 1 microgram per ml [1uL in 1mL] DAPI in H2O for 5 min. (DAPI crashes out in PBS)\nb. wash them once in PBS \n\n''8. Mount''\na. tap the coverslip on a kimwipe to let the excess PBS run off.\nb. put a drop of VectaShield onto the coverslip\nc. invert the coverslip onto a slide\nd. seal the slide with nail-polish and let it dry.\ne. take a cotton swab and some water and just clean off any salt\nf. if you can't look at slides right away, you can keep them in the cold room for up to two weeks. slides can be frozen in the -20 for longer storage (months).
__''I''__ntra__''f''__lagellar__''T''__ransport\n\n[img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_40gwdp9m99]]\n\n
[img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_4fxv95bdn][http://www.jbc.org/cgi/content/full/280/30/27688/FIG8]]\n[[Doug Cole]] [[Lucker2005|http://www.jbc.org/cgi/content/full/280/30/27688]]
Peripheral component of [[IFT Complex B]]\n\nInteracts with Chlamydomonas [[EB1]] at the flagellar tip.\n\nEncoded by ts mutant [[fla11]]
Peripheral component of [[IFT Complex B]]\n\nBinds strongly to [[KIF3B]] and [[IFT57]] linking KinesinII to the IFTB complex (mammals). [[Baker2003|http://www.jbc.org/cgi/content/abstract/278/36/34211]]\n[img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_2dhnsz2r7][http://www.jbc.org/cgi/content/full/278/36/34211/FIG8]]\n\nAssociated with the Golgi, as well as the basal body and cillium. [[IFT20-GFP]] is highly dynamic and moves between the Golgi complex and the cilium as well as along ciliary microtubules. Strong knock down of IFT20 in mammalian cells blocks ciliary assembly but does not affect Golgi structure. Moderate knockdown does not block cilia assembly but reduces the amount of polycystin-2 that is localized to the cilia. This suggests that IFT20 functions in the delivery of ciliary membrane proteins from the Golgi complex to the cilium.[[Follit2006|http://www.molbiolcell.org/cgi/content/abstract/17/9/3781]]\n\n[[Greg Pazour]]
Small Rab-like GTPase. [[Quin2007|http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1905864]]\n\n
"the full-length genomic DNA of IFT27, without its stop codon, including the 1.5 kb upstream promoter region (total 4 kb) was obtained through PCR from total genomic DNA. The full-length GFP fragment including the 230 bp downstream 30UTR (total 1.0 kb) was obtained through PCR from a CrGFP plasmid [S9]. These two PCR products were mixed and used as the third PCR template for generating the chimeric IFT27::GFP gene driven by the IFT27 native promoter. This IFT27::GFP chimeric gene fragment was cloned into a TOPO PCRII vector. Subsequently, another fragment containing the selectable marker aphVIII gene was added into the same construct."
Specifically required for transporting Outer Dynein Arms into flagella. [[Hou2007|http://www.jcb.org/cgi/content/full/176/5/653]] [[George Witman]]
[[bld1]] chlamydomonas mutant, homolog of [[OSM6]], required for flagellar assembly.\n\n[[Brazelton2001|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VRT-448Y7WK-K&_user=4430&_coverDate=10%2F16%2F2001&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=7261f48bd228578c61f4c8487b0ae608]], [[Pete LeFebvre]]\n[[Deane2001|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VRT-448Y7WK-J&_user=4430&_coverDate=10%2F16%2F2001&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=f869ebae94f360563d1a5301a22b018d]], [[Joel Rosenbaum]]\n\nProcessive retrograde in Trypanosomes (Johanna Buisson and Phelippe Bastin) and C. elegans [[Mukhopadhyay2007|http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=17510633]]
Peripheral component of [[IFT Complex B]]\n\nHelps complex B bind KinesinII / [[KIF3B]] through an interaction with [[IFT20]]\n\nIn fish photoreceptor cells, IFT20 binds particles through IFT57. They mediate the interaction with Kinesin-II and may be required for kinesin dissociation. [[Krok2008|http://www.ncbi.nlm.nih.gov/pubmed/18492793]]
[[Kobayashi2007|http://www.blackwell-synergy.com/doi/full/10.1111/j.1365-2443.2007.01076.x]] Both [[IFT81]] and IFT74 are required for [[IFT]] in worms.
[[Kobayashi2007|http://www.blackwell-synergy.com/doi/full/10.1111/j.1365-2443.2007.01076.x]] Both IFT81 and [[IFT74]] are required for [[IFT]] in worms.
First lab to make phtoactivatable GFP - mutated EGFP to WEGFP to [[PA-GFP]]. [[Patterson2002|http://www.sciencemag.org/cgi/content/full/297/5588/1873]]
The non-motor subunit of the heterotrimeric kinesin that powers antrograde [[IFT]].\nchlamydomonas mutants: [[fla3]] and [[KAP-GFP]]
GFP labeled [[KAP]] non-motor subunit of heterotrimeric kinesin.\nMade by fusing the full wt fla3 gene to chlamy GFP and integrating this transcript into the [[fla3]] mutant. [[Mueller05|http://www.molbiolcell.org/cgi/content/abstract/16/3/1341]]\n[[Joshua Mueller]] and [[Mary Porter]]\n
a.k.a mouse [[KIF3B]], One of the two motor subunits of the heterotrimeric kinesin that powes antrograde [[IFT]]\nBinds [[IFT Complex B]] through an interaction with [[IFT20]] [[Baker2003|http://www.jbc.org/cgi/content/abstract/278/36/34211]]\n\n[[fla10]] Chlamy mutant
a.k.a. mouse [[KIF3A]], One of the two motor subunits of the heterotrimeric kinesin that powes antrograde [[IFT]]\n\n[[fla8]] chlamy mutant
Mouse homolog of the two motor subunits of the heterotrimeric kinesin that powes antrograde [[IFT]]\nBinds [[IFT Complex B]] through an interaction with [[IFT20]] [[Baker2003|http://www.jbc.org/cgi/content/abstract/278/36/34211]]\n\nmutant [[KIF3B]] found that can assosciate with [[KIF3A]] but not [[KAP]] [[Haraguchi2006|http://www.jbc.org/cgi/content/abstract/281/7/4094]]
From stony coral, Trachyphyllia geoffroyi. Exhibits an irreversible spectral shift from green to red fluorescence after activation by ultraviolet light (350-410nm).\nAndo2002\nDittrich2005
Drubin endocytosis
__MT PCR__\n1) Pick chlamy off of plate and suspend in 100uL ddH2O (PCR strips)\n2) Freeze at -80\n3) Boil 10min in PCR machiene\n4) Vortex to mix, spin down 1 minute\n5) Use 5uL crude extract per 25uL GC-PCR reaction:\n\n5uL crude extract DNA (less if using purified genomic DNA)\n7uL ddH2O\n5uL Buffer GC\n5uL GC Melt\n1uL M3 Primer pair (20uL MTM1176F + 20uL MTM1865R + 60uL ddH20)\n1uL P2 Primer pair (20uL MTP1446F + 20uL MTP1869R + 60uL ddH2O)\n.5uL dNTP (10mM)\n.5uL GC Polymerase
[[Chlamy Mutants]]\n[[Proteins]]\n[[Drugs]]\n[[People]]\n[[Vocabulary]]\n[[Related Topics]]\n[[Resources]]\n[[Protocols]]
[[Mueller05|http://www.molbiolcell.org/cgi/content/abstract/16/3/1341]]
__''Media:''__\n[[TAP]]\n\n__''Buffers:''__\n[[CTAB Buffer]]\n[[1M Tris/Hcl]]\n[[5M NaCl]]\n[[10x TBS]]
Analogous transport system to [[IFT]], in that it relies on the same molecular motors: ''heterotrimeric kinesin-2'' and ''cytoplasmic dynein''. Best characterized in zebrafish melanocytes, though it also occurs in amphibians, reptiles, crustaceans, and mammals. Animals change color by altering the distribution of melanosomes (lysosome-related pigment organelles) within their melanocyte cells. Melanosome motility varies from [[IFT]], in that it also relies on myosin for short range movements on the actin cytoskeleton at the cell periphery. Defects in either the microtubule or actin motilities result in pigment diseases, such albinism.\n[img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_36qff88vgj]] [img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_38gtkqjpgd]] \n\n[[Barral2004|http://www.blackwell-synergy.com/doi/full/10.1111/j.1600-0749.2004.00138.x]], [[Gelfand2003|http://arjournals.annualreviews.org/doi/pdf/10.1146/annurev.cellbio.19.111401.092937]], [[Skold2002|http://www3.interscience.wiley.com/cgi-bin/fulltext/98517527/PDFSTART]]\n\nMotor cooperation in Melanosome transport [[Levi2006|http://www.biophysj.org/cgi/content/full/90/1/318]]\n\nDynactin may mediate the switching between antrograde and retrograde transport (competition between dynein and kinesin-2 binding) in melanosomes [[JCBnewsviews2003|http://www.jcb.org/cgi/content/full/160/3/291]], [[Deacon2003|http://www.jcb.org/cgi/content/full/160/3/297?ijkey=cf363d60ec3d47d0b992ae6d531db84868ce2683]]
WEGFP histadine for threonine substitution at position 203 (T203H) reduces minor peak absorbance prior to ~400nm photoactivation. \n\n"To facilitate their use in mammalian cell biology experiments, we generated mutations in a codon-optimized GFP variant developed from the commercially available EGFP. We made __''L64F''__ and __''T65S''__ mutations within EGFP to recover wild-type GFP absorbance properties consisting of a major peak at 397 nm and a minor peak at 475 nm. An additional __''V163A''__ mutation was made to improve folding at 37°C. This construct is referred to as WEGFP. Several T203 mutations, in addition to T203I, were made in this construct which reduced the minor peak and retained the major peak at ~400 nm (Table S1). While many mutants had spectral properties that were comparable with the T203I, the __''T203H''__ mutant was exceptional and decreased the minor absorbance peak even more drastically than the isoleucine mutation. The T203H mutant proved to be an excellent photoactivatable marker and was named [[PA-GFP]]." \n[img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_8c45z5zc7][http://www.sciencemag.org/cgi/content/full/297/5588/1873/F1]][img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_10xzjh5qzp]]\n[[Patterson2002|http://www.sciencemag.org/cgi/content/full/297/5588/1873]], [[Jennifer Lippincott-Schwartz]]\n
Ahmet's old boss. Invented [[FIONA]] for tracking motor movements in vitro, leading to Ahmet's observation that Kinesin walks hand over hand (like Myosin). \n[[Yildiz2004|http://www.sciencemag.org/cgi/content/full/303/5658/676]]\n[[Yildiz2003|http://www.sciencemag.org/cgi/content/full/300/5628/2061?ijkey=5681642aa8c20537e28cc0a49f415d903912fb50]]\n\nHe later used [[FIONA]] in vivo to show that multiple dyneins or kinesins can push a peroxysome in unison, achieving speeds up to 10x faster than in vitro (up to ''12 um/sec'')\n[img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_16cq88q3hm]]\n[[Kural2005|http://www.sciencemag.org/cgi/content/full/308/5727/1469]]
__''Chlamy / Flagellar Length Control:''__\n[[Mike Adams]]\n[[Bob Bloodgood]]\n[[Doug Cole]]\n[[William Dentler]]\n[[Dennis Diener]]\n[[Patrick Ferris]]\n[[Stefan Geimer]]\n[[Ursula Goodenough]]\n[[Elizabeth Harris]]\n[[Jonathan Jarvik]]\n[[Keith Kozminski]]\n[[Pete LeFebvre]]\n[[David Luck]]\n[[Wallace Marshall]]\n[[Greg Pazour]]\n[[Lotte Pedersen]]\n[[Gianni Piperno]]\n[[Mary Porter]]\n[[Lynne Quarmby]]\n[[Joel Rosenbaum]]\n[[Roger Sloboda]]\n[[William Snell]]\n[[Nedra Wilson]]\n[[George Witman]]\n\n__''Microscopy:''__\n[[Mats Gustafsson]]\n[[Jennifer Lippincott-Schwartz]]\n[[Paul Selvin]]\n[[Roger Tsien]]\n[[Clare Waterman-Storer]]\n[[Torsten Wittmann]]\n
__Chlamy__\n[[Traffiking Flagellar Proteins]]\n[[Structural Flagellar Proteins]]\n[[Basal Body Proteins]]\n[[Transition Zone Proteins]]\n\n__Microscopy__\n[[Fluorescent Proteins]]
[[Chlamy Transformation]]\n[[Chlamy Mating]]\n[[IF Staining - Chlamy]]\n[[Flagellar Prep]]\n[[Genomic DNA Prep]]\n[[pH Shock]]\n[[Linkage Mapping and MT PCR]]\n[[PCR Programs]]\n[[Bradford Assay]]\n[[Media and Buffers]]
[[Waters2005|http://arjournals.annualreviews.org/doi/full/10.1146/annurev.cellbio.21.012704.131001]]
Motor Interactions Reviews [[Mallik2004|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VRT-4DVM8BN-J&_user=4430&_coverDate=11%2F23%2F2004&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=9c5d5fd3c10aaebbb9e7413c53521048]], [[Vale2003|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WSN-4811J61-6&_user=4430&_coverDate=02%2F21%2F2003&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=95b0e6aec0a2d81fbf53e37f55ff2356]], [[\n\nKinesin Family Overview [[Miki2005|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TCX-4GV2NDV-1&_user=4430&_coverDate=09%2F30%2F2005&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=ae52abc0ce4dfb182eebdbee73028d69]]\n\nKinesin-1 inactivates itself when not bound to cargo: the cargo-binding C-terminal tail interacts with the N-terminal motor domains [[FriedmanVale99|http://www.nature.com/ncb/journal/v1/n5/full/ncb0999_293.html]], [[Stock99|http://www.jbc.org/cgi/content/full/274/21/14617]], [[Verhey98|http://www.jcb.org/cgi/content/full/143/4/1053]]\nKinesin-1 is activated in two steps [[JCBnewsviews2007|http://www.jcb.org/cgi/content/full/176/1/2]], [[Hackney2007|http://www.jcb.org/cgi/content/full/176/1/7]], [[Cai2007|http://www.jcb.org/cgi/content/full/176/1/51]], [[Blasius2007|http://www.jcb.org/cgi/content/full/176/1/11]]\n[[Endow2003|http://arjournals.annualreviews.org/doi/pdf/10.1146/annurev.physiol.65.092101.142550]]\n\nKinesin-1 may DIRECTLY bind and transport Cytoplasmic Dynein (interaction between DIC and KLC). [[Ligon2004|[http://www.jbc.org/cgi/content/full/279/18/19201]]\n\nDynactin increases Dynein processivity [[King1999|http://www.nature.com/ncb/journal/v2/n1/full/ncb0100_20.html]] and is required for [[Axonal Transport]] [[Lamonte2002|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WSS-4621C50-10&_user=4430&_coverDate=05%2F30%2F2002&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=c5fc93c40744026fd42e0b3d0ee8ef4a]], [[Schroer2004|http://arjournals.annualreviews.org/doi/full/10.1146/annurev.cellbio.20.012103.094623]], [[Levy2005|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T01-4J021P2-1&_user=4430&_coverDate=05%2F31%2F2006&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=d756d0441787c8233c1270be5aa21fb4#SECX4]]\nDynactin may mediate the switching between antrograde and retrograde transport (competition between dynein and kinesin-2 binding) in melanosomes [[JCBnewsviews2003|http://www.jcb.org/cgi/content/full/160/3/291]], [[Deacon2003|http://www.jcb.org/cgi/content/full/160/3/297?ijkey=cf363d60ec3d47d0b992ae6d531db84868ce2683]]\nDynamin P150glued interacts w/ EB1 and CLIP-170 at the +end, but tracks with transporting particles [[Watson2006|http://jcs.biologists.org/cgi/reprint/119/13/2758]]\n''Is there Dynactin in Flagella?''\n\nPlus-end tracking proteins regulate Dynein cargo loading (viat Dynactin interactions) [[Wu2006|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TCX-4J8KVXP-1&_user=4430&_coverDate=03%2F31%2F2006&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=3ef591128962b58d3f0f3c103141d49f]]\n\n\n
__''Techniques''__\n[[Kymograph advantages and limitations]]\n[[Fourier Transform advantages and limitations]]\n[[Photoactivation advantages and limitations]]\n\n__''Concepts and Background''__\n[[Diffusion]]\n[[Periodicity]]\n[[Small GTPases]]\n[[+TIPs]]\n[[Regulation of Kinesin and Dynein Activity]]\n[[Ciliary Diseases]]\n\n__''Analogous Systems''__\n[[Axonal Transport]]\n[[Melanosome Motility]]\n[[Bacterial Flagellar Hook Length Control]]\n[[Bacteriophage Tail Length Control]]\n[[Spindle Size Control]]\n[[Cell Size Control]]\n[[Organ Size Control]]\n[[Quorum Sensing]]\n[[Developmental Spacing]]\n[[Biological Gates]]\n[[Biological Oscillators]]\n
[[Chlamy Codon Usage]]\n\n[[Chlamy Center|http://www.chlamy.org/]]\n\n[[Chlamy Flagellar Proteome|http://labs.umassmed.edu/chlamyfp/index.php]]\n\n[[Chlamy Message Board|http://www.bio.net/bionet/mm/chlamy/]]\n\n[[Chlamy FAQ|http://www.yale.edu/rosenbaum/green_yeast.html]]\n\n[[Chlamy Labs|http://www.yale.edu/rosenbaum/chlamylist.html]]
Striated Fiber Assemblin, major component of the striated microtubule-associated fibers at the basal body (run along rootlets)\n[[Lechtreck2002|http://jcs.biologists.org/cgi/content/full/115/7/1511]]
__''S''__ite __''o''__f __''F''__lagellar __''A''__utonomy\n\nThis is the region just distal of the [[transition zone]] where the flagella is cleaved following [[pH Shock]]. Cleavage is thought to be mediated by [[fa1]] / [[fa2]] and [[katanin]].
flagellar length control and info on eveyone's favorite green algae
Chlamydomonas
Rab functions in primary cilia [[Yoshimura2007|http://www.jcb.org/cgi/content/full/178/3/363]]\n\nG-protein control of MT assembly [[Zheng2004|http://arjournals.annualreviews.org/doi/full/10.1146/annurev.cellbio.20.012103.094648]]\n\nRho biochemistry and biology [[Hall2005|http://arjournals.annualreviews.org/doi/full/10.1146/annurev.cellbio.21.020604.150721]]\nArfs in transport [[Chavrier2006|http://www.nature.com/nrm/journal/v7/n5/full/nrm1910.html]]\nhow Arls differ from Arfs [[Burd2004|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TCX-4DTM0HW-1&_user=4430&_coverDate=12%2F01%2F2004&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=c45e69b0433b795551a9afc25f06e47f]]\n\nRabs and Traffic, Rab27 needed for myosin interaction in [[Melanosome Motility]] [[Novick2006|http://www.pnas.org/cgi/content/full/103/32/11821]]\nRabs on endosomal membranes [[Deneka2002|http://www.nature.com/ncb/journal/v4/n2/full/ncb0202-e33.html]] Rab family [[Stenmark2001|http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=11387043]]\n\nPlant small GTPases and vesicle transport [[Molendijk2004|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VS4-4DD9GF3-5&_user=4430&_coverDate=12%2F01%2F2004&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=a2719915c9aac0788e4995270d39d1e0]]
Spindel Length is determined by microtubule dynamics. Microtubules in the spindle treadmill- more addition/stabilization = longer, more depolymerization = shorter.\n[[Goshima2005|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VRT-4HM7R39-N&_user=4430&_coverDate=11%2F22%2F2005&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=c423a025dff3e4c0c5620ce37c8905e8]], [[Gaetz2004|http://www.jcb.org/cgi/content/full/166/4/465]]\n\nDynein/Dynacin transport of NuMA to -end of microtubules is required for spindle assembly. [[Merdes2000|http://www.jcb.org/cgi/content/full/149/4/851]]\n\nmore speckling:\n+ end dynamics [[Tirnaeur2004|http://www.molbiolcell.org/cgi/content/full/15/4/1776]], - end distributions throughout spindles [[Burbank2006|http://www.jcb.org/cgi/content/full/175/3/369]]
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{\n background: [[ColorPalette::PrimaryLight]];\n border: 1px solid [[ColorPalette::PrimaryMid]];\n}\n\n.popup hr {\n color: [[ColorPalette::PrimaryDark]];\n background: [[ColorPalette::PrimaryDark]];\n border-bottom: 1px;\n}\n\n.listBreak div{\n border-bottom: 1px solid [[ColorPalette::PrimaryDark]];\n}\n\n.popup li.disabled {\n color: [[ColorPalette::PrimaryDark]];\n}\n\n.popup li a, .popup li a:visited {\n color: [[ColorPalette::PrimaryMid]];\n border: none;\n}\n\n.popup li a:hover {\n background: [[ColorPalette::PrimaryDark]];\n color: [[ColorPalette::Background]];\n border: none;\n}\n\n.tiddler .defaultCommand {\n font-weight: bold;\n}\n\n.shadow .title {\n color: [[ColorPalette::TertiaryDark]];\n}\n\n.title {\n color: [[ColorPalette::SecondaryDark]];\n}\n\n.subtitle {\n color: [[ColorPalette::TertiaryDark]];\n}\n\n.toolbar {\n color: [[ColorPalette::PrimaryMid]];\n}\n\n.tagging, .tagged {\n border: 1px solid [[ColorPalette::TertiaryPale]];\n background-color: [[ColorPalette::TertiaryPale]];\n}\n\n.selected .tagging, .selected .tagged {\n background-color: [[ColorPalette::TertiaryLight]];\n border: 1px solid [[ColorPalette::TertiaryMid]];\n}\n\n.tagging .listTitle, .tagged .listTitle {\n color: [[ColorPalette::PrimaryDark]];\n}\n\n.tagging .button, .tagged .button {\n border: none;\n}\n\n.footer {\n color: [[ColorPalette::TertiaryLight]];\n}\n\n.selected .footer {\n color: [[ColorPalette::TertiaryMid]];\n}\n\n.sparkline {\n background: [[ColorPalette::PrimaryPale]];\n border: 0;\n}\n\n.sparktick {\n background: [[ColorPalette::PrimaryDark]];\n}\n\n.error, .errorButton {\n color: [[ColorPalette::Foreground]];\n background: [[ColorPalette::Error]];\n}\n\n.warning {\n color: [[ColorPalette::Foreground]];\n background: [[ColorPalette::SecondaryPale]];\n}\n\n.cascade {\n background: [[ColorPalette::TertiaryPale]];\n color: [[ColorPalette::TertiaryMid]];\n border: 1px solid [[ColorPalette::TertiaryMid]];\n}\n\n.imageLink, #displayArea .imageLink {\n background: transparent;\n}\n\n.viewer .listTitle {list-style-type: none; margin-left: -2em;}\n\n.viewer .button {\n border: 1px solid [[ColorPalette::SecondaryMid]];\n}\n\n.viewer blockquote {\n border-left: 3px solid [[ColorPalette::TertiaryDark]];\n}\n\n.viewer table {\n border: 2px solid [[ColorPalette::TertiaryDark]];\n}\n\n.viewer th, thead td {\n background: [[ColorPalette::SecondaryMid]];\n border: 1px solid [[ColorPalette::TertiaryDark]];\n color: [[ColorPalette::Background]];\n}\n\n.viewer td, .viewer tr {\n border: 1px solid [[ColorPalette::TertiaryDark]];\n}\n\n.viewer pre {\n border: 1px solid [[ColorPalette::SecondaryLight]];\n background: [[ColorPalette::SecondaryPale]];\n}\n\n.viewer code {\n color: [[ColorPalette::SecondaryDark]];\n}\n\n.viewer hr {\n border: 0;\n border-top: dashed 1px [[ColorPalette::TertiaryDark]];\n color: [[ColorPalette::TertiaryDark]];\n}\n\n.highlight, .marked {\n background: [[ColorPalette::SecondaryLight]];\n}\n\n.editor input {\n border: 1px solid [[ColorPalette::PrimaryMid]];\n}\n\n.editor textarea {\n border: 1px solid [[ColorPalette::PrimaryMid]];\n width: 100%;\n}\n\n.editorFooter {\n color: [[ColorPalette::TertiaryMid]];\n}\n\n/*}}}*/
https://www.chlamycollection.org/methods/media-recipes/tap-and-tris-minimal/\n\n\n__''1000x Phosphate Buffer II (for 100 ml)''__\n\nK2HPO4 10.8 g\n\nKH2PO4 5.6 g\n\n \n\n__''100x TAP Salts (for 500 ml)''__ (a.k.a. Solution A)\n\n\nNH4Cl 20 g\n\nMgSO4-7H2O 5 g\n\nCaCl2-2H2O 2.5 g\n\n\n\n__''TAP liquid media''__\n\n1. Put ~800 ml deionized water into a beaker, flask, or bottle.\n\n2. Add the following solutions:\n\na. 10 ml 100x tris buffer\n\nb. 10 ml 100x TAP salts\n\nc. 1 ml 1000x phosphate buffer\n\nd. 1 ml Hutner trace elements\n\ne. 1 ml glacial acetic acid\n\n3. Add deionized water to a total volume of 1 L.\n\n4. Test the pH of the solution. If it is above 7, add more acetic acid to adjust the pH. If the pH is less than 7, throw the solution away and start again.\n\n5. Autoclave 40 minutes on liquid cycle.\n\n\n__''TAP + arg liquid media''__\n\n6. After the media has cooled, add 10ml of 100x arginine solution (kept in the freezer).\n\n\n__''TAP -N media"__\n\nOmit the acetic acid and titrate the final solution to pH 7.0 with HCl\n\n\n__''TAP agar plates''__\n
__T__otal __I__nternal __R__eflection __F__luorescence Microscopy
__Heterotrimeric Kinesin:__\n[[KHP1]] ([[fla10]] / [[fla7]]) - kinesin motor subunit\n"[[KHP2]]" ([[fla8]] / [[fla1]]) - kinesin motor subunit\n[[KAP]] ([[fla3]]) - kinesin non-motor subunit\n\n__Cytoplasmic Dynein:__\n[[DHC1b]] - dynein heavy chain\n[[D1bLIC]] - dynein intermediate light chain\n[[LC8]] - light chain shared by several proteins\n\n__[[IFT Complex B]]:__ (15-16S, 710-760kDa) \n[[IFT88]] - core IFTB subunit\n[[IFT81]] - core IFTB subunit\n[[IFT74]] - core IFTB subunit\n[[IFT72]] - core IFTB subunit\n[[IFT52]] - core IFTB subunit\n[[IFT46]] - core IFTB subunit\n[[IFT27]] - core IFTB subunit\n[[IFT172]] - peripheral IFTB subunit\n[[IFT80]] - peripheral IFTB subunit\n[[IFT57]] - peripheral IFTB subunit\n[[IFT20]] - peripheral IFTB subunit\n\n__IFT Complex A__ (16S, 550kDa)\n[[IFT144]]\n[[IFT140]]\n[[IFT139]]\n[[IFT122]]\n\n[[Doug Cole Analysis of IFT Complex Composition|http://www.jcb.org/cgi/content/full/141/4/993/TIII]]\n\n__OTHERS (capping, length control):__\n[[EB1]] - MT +end binder\n[[GSK3]]\n[[CLASPs]]
Fa1p\nFa2p\nKatanin\nCnk2p\nNek8
/***\nContains the stuff you need to use Tiddlyspot\nNote you must also have UploadPlugin installed\n***/\n//{{{\n\n// edit this if you are migrating sites or retrofitting an existing TW\nconfig.tiddlyspotSiteId = 'chlamy';\n\n// make it so you can by default see edit controls via http\nconfig.options.chkHttpReadOnly = false;\n\n// disable autosave in d3\nif (window.location.protocol != "file:")\n config.options.chkGTDLazyAutoSave = false;\n\n// tweak shadow tiddlers to add upload button, password entry box etc\nwith (config.shadowTiddlers) {\n SiteUrl = 'http://'+config.tiddlyspotSiteId+'.tiddlyspot.com';\n SideBarOptions = SideBarOptions.replace(/(<<saveChanges>>)/,"$1<<tiddler TspotSidebar>>");\n OptionsPanel = OptionsPanel.replace(/^/,"<<tiddler TspotOptions>>");\n DefaultTiddlers = DefaultTiddlers.replace(/^/,"[[Welcome to Tiddlyspot]] ");\n MainMenu = MainMenu.replace(/^/,"[[Welcome to Tiddlyspot]] ");\n}\n\n// create some shadow tiddler content\nmerge(config.shadowTiddlers,{\n\n'Welcome to Tiddlyspot':[\n "This document is a ~TiddlyWiki from tiddlyspot.com. A ~TiddlyWiki is an electronic notebook that is great for managing todo lists, personal information, and all sorts of things.",\n "",\n "@@font-weight:bold;font-size:1.3em;color:#444; //What now?// &nbsp;&nbsp;@@ Before you can save any changes, you need to enter your password in the form below. Then configure privacy and other site settings at your [[control panel|http://" + config.tiddlyspotSiteId + ".tiddlyspot.com/controlpanel]] (your control panel username is //" + config.tiddlyspotSiteId + "//).",\n "<<tiddler TspotControls>>",\n "See also GettingStarted.",\n "",\n "@@font-weight:bold;font-size:1.3em;color:#444; //Working online// &nbsp;&nbsp;@@ You can edit this ~TiddlyWiki right now, and save your changes using the \s"save to web\s" button in the column on the right.",\n "",\n "@@font-weight:bold;font-size:1.3em;color:#444; //Working offline// &nbsp;&nbsp;@@ A fully functioning copy of this ~TiddlyWiki can be saved onto your hard drive or USB stick. You can make changes and save them locally without being connected to the Internet. When you're ready to sync up again, just click \s"upload\s" and your ~TiddlyWiki will be saved back to tiddlyspot.com.",\n "",\n "@@font-weight:bold;font-size:1.3em;color:#444; //Help!// &nbsp;&nbsp;@@ Find out more about ~TiddlyWiki at [[TiddlyWiki.com|http://tiddlywiki.com]]. Also visit [[TiddlyWiki Guides|http://tiddlywikiguides.org]] for documentation on learning and using ~TiddlyWiki. New users are especially welcome on the [[TiddlyWiki mailing list|http://groups.google.com/group/TiddlyWiki]], which is an excellent place to ask questions and get help. If you have a tiddlyspot related problem email [[tiddlyspot support|mailto:support@tiddlyspot.com]].",\n "",\n "@@font-weight:bold;font-size:1.3em;color:#444; //Enjoy :)// &nbsp;&nbsp;@@ We hope you like using your tiddlyspot.com site. Please email [[feedback@tiddlyspot.com|mailto:feedback@tiddlyspot.com]] with any comments or suggestions."\n].join("\sn"),\n\n'TspotControls':[\n "| tiddlyspot password:|<<option pasUploadPassword>>|",\n "| site management:|<<upload http://" + config.tiddlyspotSiteId + ".tiddlyspot.com/store.cgi index.html . . " + config.tiddlyspotSiteId + ">>//(requires tiddlyspot password)//<<br>>[[control panel|http://" + config.tiddlyspotSiteId + ".tiddlyspot.com/controlpanel]], [[download (go offline)|http://" + config.tiddlyspotSiteId + ".tiddlyspot.com/download]]|",\n "| links:|[[tiddlyspot.com|http://tiddlyspot.com/]], [[FAQs|http://faq.tiddlyspot.com/]], [[announcements|http://announce.tiddlyspot.com/]], [[blog|http://tiddlyspot.com/blog/]], email [[support|mailto:support@tiddlyspot.com]] & [[feedback|mailto:feedback@tiddlyspot.com]], [[donate|http://tiddlyspot.com/?page=donate]]|"\n].join("\sn"),\n\n'TspotSidebar':[\n "<<upload http://" + config.tiddlyspotSiteId + ".tiddlyspot.com/store.cgi index.html . . " + config.tiddlyspotSiteId + ">><html><a href='http://" + config.tiddlyspotSiteId + ".tiddlyspot.com/download' class='button'>download</a></html>"\n].join("\sn"),\n\n'TspotOptions':[\n "tiddlyspot password:",\n "<<option pasUploadPassword>>",\n ""\n].join("\sn")\n\n});\n//}}}\n
| !date | !user | !location | !storeUrl | !uploadDir | !toFilename | !backupdir | !origin |\n| 1/6/2007 13:19:31 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 13:20:57 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 13:28:32 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 13:42:39 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 13:43:44 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 13:45:7 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 1/6/2007 13:52:47 | Bengel | [[/|http://chlamy.tiddlyspot.com/#StyleSheetColors]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 14:30:34 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 1/6/2007 19:49:27 | Bengel | [[/|http://chlamy.tiddlyspot.com/#StyleSheetColors]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 19:54:52 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5B1%20June%202007%5D%5D%20%5B%5BNew%20Tiddler%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 21:29:3 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5B1%20June%202007%5D%5D%20%5B%5BNew%20Tiddler%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 21:41:50 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5B1%20June%202007%5D%5D%20%5B%5BNew%20Tiddler%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 1/6/2007 21:42:25 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5B1%20June%202007%5D%5D%20%5B%5BNew%20Tiddler%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 1/6/2007 21:43:2 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5B1%20June%202007%5D%5D%20%5B%5BNew%20Tiddler%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 22:3:42 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5B1%20June%202007%5D%5D%20%5B%5BNew%20Tiddler%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 22:17:14 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5B1%20June%202007%5D%5D%20%5B%5BNew%20Tiddler%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 22:31:39 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5B1%20June%202007%5D%5D%20%5B%5BNew%20Tiddler%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 23:15:14 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5B1%20June%202007%5D%5D%20%5B%5BNew%20Tiddler%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 23:30:45 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5B1%20June%202007%5D%5D%20%5B%5BNew%20Tiddler%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 1/6/2007 23:59:4 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5B1%20June%202007%5D%5D%20%5B%5BNew%20Tiddler%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 2/6/2007 0:13:43 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5B1%20June%202007%5D%5D%20%5B%5BNew%20Tiddler%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 2/6/2007 0:34:1 | Bengel | [[/|http://chlamy.tiddlyspot.com/#pf1]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 2/6/2007 0:40:14 | Bengel | [[/|http://chlamy.tiddlyspot.com/#StyleSheetColors]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 2/6/2007 0:46:35 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 2/6/2007 0:55:29 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 2/6/2007 0:56:32 | Bengel | [[index.html|http://chlamy.tiddlyspot.com/index.html]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 2/6/2007 1:6:20 | Bengel | [[index.html|http://chlamy.tiddlyspot.com/index.html#StyleSheetColors]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 2/6/2007 1:7:23 | Bengel | [[index.html|http://chlamy.tiddlyspot.com/index.html#StyleSheetColors]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 2/6/2007 1:16:20 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 2/6/2007 1:17:43 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 8/6/2007 11:32:49 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 8/6/2007 12:14:52 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 8/6/2007 12:43:18 | Bengel | [[/|http://chlamy.tiddlyspot.com/#IFT20]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 8/6/2007 12:53:6 | Bengel | [[/|http://chlamy.tiddlyspot.com/#IFT20]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 8/6/2007 14:59:15 | Bengel | [[/|http://chlamy.tiddlyspot.com/#IFT20]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 8/6/2007 15:41:47 | Bengel | [[/|http://chlamy.tiddlyspot.com/#IFT20]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 8/6/2007 19:38:10 | Bengel | [[/|http://chlamy.tiddlyspot.com/#IFT20]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 18/6/2007 16:33:49 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 18/6/2007 17:0:21 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 18/6/2007 17:32:18 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 19/6/2007 13:0:48 | Bengel | [[index.20070619.1243090000.html|http://chlamy.tiddlyspot.com/backup/index.20070619.1243090000.html]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 19/6/2007 13:24:54 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 19/6/2007 13:26:57 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 19/6/2007 15:0:26 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 19/6/2007 15:10:33 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 19/6/2007 15:20:33 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 20/6/2007 19:31:5 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 20/6/2007 19:32:34 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 21/6/2007 15:19:33 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5Bphotoactivation%20%2F%20photoconversion%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 21/6/2007 15:54:48 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5Bphotoactivation%20%2F%20photoconversion%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 21/6/2007 16:40:56 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5Bphotoactivation%20%2F%20photoconversion%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 21/6/2007 17:9:50 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 21/6/2007 17:14:31 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 21/6/2007 18:7:8 | Bengel | [[/|http://chlamy.tiddlyspot.com/#cGFP]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 21/6/2007 18:37:21 | Bengel | [[/|http://chlamy.tiddlyspot.com/#cGFP]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 21/6/2007 19:2:14 | Bengel | [[/|http://chlamy.tiddlyspot.com/#cGFP]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 21/6/2007 19:5:47 | Bengel | [[/|http://chlamy.tiddlyspot.com/#cGFP]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 21/6/2007 19:22:19 | Bengel | [[/|http://chlamy.tiddlyspot.com/#cGFP]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 21/6/2007 19:22:48 | Bengel | [[/|http://chlamy.tiddlyspot.com/#cGFP]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 21/6/2007 19:32:30 | Bengel | [[/|http://chlamy.tiddlyspot.com/#cGFP]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 21/6/2007 19:38:54 | Bengel | [[/|http://chlamy.tiddlyspot.com/#cGFP]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 21/6/2007 19:39:16 | Bengel | [[/|http://chlamy.tiddlyspot.com/#cGFP]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 21/6/2007 19:43:0 | Bengel | [[/|http://chlamy.tiddlyspot.com/#cGFP]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 22/6/2007 14:6:37 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 22/6/2007 16:18:20 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 22/6/2007 16:53:25 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 27/6/2007 1:16:14 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 27/6/2007 14:49:21 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 28/6/2007 2:29:47 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 28/6/2007 16:36:37 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 28/6/2007 16:38:57 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 28/6/2007 17:47:28 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 28/6/2007 17:48:41 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 28/6/2007 18:19:41 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 28/6/2007 18:43:12 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 2/7/2007 18:6:13 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 2/7/2007 18:8:50 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 2/7/2007 18:19:1 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 2/7/2007 19:52:28 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 2/7/2007 19:52:48 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 9/7/2007 15:2:24 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 9/7/2007 15:53:35 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 9/7/2007 15:56:42 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 15/7/2007 18:0:14 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/7/2007 18:48:18 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/7/2007 18:58:21 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/7/2007 19:38:47 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/7/2007 19:47:43 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 16/7/2007 14:29:54 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 16/7/2007 19:14:32 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 16/7/2007 19:15:32 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 16/7/2007 19:40:2 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 16/7/2007 22:16:38 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 16/7/2007 22:51:11 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 17/7/2007 0:29:48 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 17/7/2007 1:2:14 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 17/7/2007 1:8:5 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 17/7/2007 1:11:40 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 17/7/2007 1:25:22 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 17/7/2007 1:34:48 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 17/7/2007 1:41:29 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 17/7/2007 1:55:52 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 17/7/2007 11:42:35 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 17/7/2007 11:45:28 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 17/7/2007 12:11:27 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 17/7/2007 15:21:16 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 18/7/2007 11:11:57 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 18/7/2007 12:19:16 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 18/7/2007 13:42:7 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 18/7/2007 14:14:31 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 18/7/2007 14:48:14 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 18/7/2007 15:9:16 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 18/7/2007 17:30:51 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 11/8/2007 16:9:39 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 13/8/2007 7:46:40 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 14/8/2007 14:29:44 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 14/8/2007 14:49:58 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 14/8/2007 17:31:44 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 15/8/2007 15:38:22 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/8/2007 15:42:23 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/8/2007 15:44:46 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/8/2007 16:34:18 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/8/2007 17:18:17 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/8/2007 17:24:30 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/8/2007 21:45:0 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/8/2007 21:48:45 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/8/2007 21:53:40 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/8/2007 22:3:7 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5BGFP%20tagged%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/8/2007 22:40:53 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5BGFP%20tagged%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/8/2007 22:43:15 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5BGFP%20tagged%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 15/8/2007 23:3:59 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5BGFP%20tagged%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 16/8/2007 12:8:26 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5BGFP%20tagged%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 18/10/2007 17:18:15 | YourName | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 22/10/2007 14:57:49 | YourName | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 25/10/2007 9:23:45 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 25/10/2007 9:55:57 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 25/10/2007 9:56:59 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 25/10/2007 17:39:3 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 25/10/2007 18:59:42 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 26/10/2007 13:15:21 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 26/10/2007 15:38:31 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 7/3/2008 19:20:50 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 1/4/2008 11:32:10 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 1/4/2008 11:32:30 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 1/4/2008 11:38:59 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 1/4/2008 13:56:24 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 1/4/2008 13:56:27 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok | Ok |\n| 1/4/2008 15:48:17 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 30/4/2008 12:2:24 | Bengel | [[/|http://chlamy.tiddlyspot.com/#%5B%5BIF%20Staining%20-%20Chlamy%5D%5D]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 25/11/2008 10:44:39 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 25/11/2008 11:27:42 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 2/2/2009 13:39:13 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 2/2/2009 13:40:44 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 2/2/2009 13:47:1 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 2/2/2009 14:5:4 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 2/2/2009 20:26:48 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 4/11/2009 11:40:19 | bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 4/11/2009 11:40:25 | bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 14/11/2017 10:9:58 | Ben | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 14/11/2017 10:10:18 | Ben | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 14/11/2017 10:13:0 | Ben | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 14/11/2017 10:13:7 | Ben | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 14/11/2017 10:15:22 | Ben | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |\n| 14/11/2017 10:15:36 | Chlamy | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . | Ok |\n| 14/11/2017 10:16:45 | Bengel | [[/|http://chlamy.tiddlyspot.com/]] | [[store.cgi|http://chlamy.tiddlyspot.com/store.cgi]] | . | index.html | . |
/***\n|''Name:''|UploadPlugin|\n|''Description:''|Save to web a TiddlyWiki|\n|''Version:''|3.4.5|\n|''Date:''|Oct 15, 2006|\n|''Source:''|http://tiddlywiki.bidix.info/#UploadPlugin|\n|''Documentation:''|http://tiddlywiki.bidix.info/#UploadDoc|\n|''Author:''|BidiX (BidiX (at) bidix (dot) info)|\n|''License:''|[[BSD open source license|http://tiddlywiki.bidix.info/#%5B%5BBSD%20open%20source%20license%5D%5D ]]|\n|''~CoreVersion:''|2.0.0|\n|''Browser:''|Firefox 1.5; InternetExplorer 6.0; Safari|\n|''Include:''|config.lib.file; config.lib.log; config.lib.options; PasswordTweak|\n|''Require:''|[[UploadService|http://tiddlywiki.bidix.info/#UploadService]]|\n***/\n//{{{\nversion.extensions.UploadPlugin = {\n major: 3, minor: 4, revision: 5, \n date: new Date(2006,9,15),\n source: 'http://tiddlywiki.bidix.info/#UploadPlugin',\n documentation: 'http://tiddlywiki.bidix.info/#UploadDoc',\n author: 'BidiX (BidiX (at) bidix (dot) info',\n license: '[[BSD open source license|http://tiddlywiki.bidix.info/#%5B%5BBSD%20open%20source%20license%5D%5D]]',\n coreVersion: '2.0.0',\n browser: 'Firefox 1.5; InternetExplorer 6.0; Safari'\n};\n//}}}\n\n////+++!![config.lib.file]\n\n//{{{\nif (!config.lib) config.lib = {};\nif (!config.lib.file) config.lib.file= {\n author: 'BidiX',\n version: {major: 0, minor: 1, revision: 0}, \n date: new Date(2006,3,9)\n};\nconfig.lib.file.dirname = function (filePath) {\n var lastpos;\n if ((lastpos = filePath.lastIndexOf("/")) != -1) {\n return filePath.substring(0, lastpos);\n } else {\n return filePath.substring(0, filePath.lastIndexOf("\s\s"));\n }\n};\nconfig.lib.file.basename = function (filePath) {\n var lastpos;\n if ((lastpos = filePath.lastIndexOf("#")) != -1) \n filePath = filePath.substring(0, lastpos);\n if ((lastpos = filePath.lastIndexOf("/")) != -1) {\n return filePath.substring(lastpos + 1);\n } else\n return filePath.substring(filePath.lastIndexOf("\s\s")+1);\n};\nwindow.basename = function() {return "@@deprecated@@";};\n//}}}\n////===\n\n////+++!![config.lib.log]\n\n//{{{\nif (!config.lib) config.lib = {};\nif (!config.lib.log) config.lib.log= {\n author: 'BidiX',\n version: {major: 0, minor: 1, revision: 1}, \n date: new Date(2006,8,19)\n};\nconfig.lib.Log = function(tiddlerTitle, logHeader) {\n if (version.major < 2)\n this.tiddler = store.tiddlers[tiddlerTitle];\n else\n this.tiddler = store.getTiddler(tiddlerTitle);\n if (!this.tiddler) {\n this.tiddler = new Tiddler();\n this.tiddler.title = tiddlerTitle;\n this.tiddler.text = "| !date | !user | !location |" + logHeader;\n this.tiddler.created = new Date();\n this.tiddler.modifier = config.options.txtUserName;\n this.tiddler.modified = new Date();\n if (version.major < 2)\n store.tiddlers[tiddlerTitle] = this.tiddler;\n else\n store.addTiddler(this.tiddler);\n }\n return this;\n};\n\nconfig.lib.Log.prototype.newLine = function (line) {\n var now = new Date();\n var newText = "| ";\n newText += now.getDate()+"/"+(now.getMonth()+1)+"/"+now.getFullYear() + " ";\n newText += now.getHours()+":"+now.getMinutes()+":"+now.getSeconds()+" | ";\n newText += config.options.txtUserName + " | ";\n var location = document.location.toString();\n var filename = config.lib.file.basename(location);\n if (!filename) filename = '/';\n newText += "[["+filename+"|"+location + "]] |";\n this.tiddler.text = this.tiddler.text + "\sn" + newText;\n this.addToLine(line);\n};\n\nconfig.lib.Log.prototype.addToLine = function (text) {\n this.tiddler.text = this.tiddler.text + text;\n this.tiddler.modifier = config.options.txtUserName;\n this.tiddler.modified = new Date();\n if (version.major < 2)\n store.tiddlers[this.tiddler.tittle] = this.tiddler;\n else {\n store.addTiddler(this.tiddler);\n story.refreshTiddler(this.tiddler.title);\n store.notify(this.tiddler.title, true);\n }\n if (version.major < 2)\n store.notifyAll(); \n};\n//}}}\n////===\n\n////+++!![config.lib.options]\n\n//{{{\nif (!config.lib) config.lib = {};\nif (!config.lib.options) config.lib.options = {\n author: 'BidiX',\n version: {major: 0, minor: 1, revision: 0}, \n date: new Date(2006,3,9)\n};\n\nconfig.lib.options.init = function (name, defaultValue) {\n if (!config.options[name]) {\n config.options[name] = defaultValue;\n saveOptionCookie(name);\n }\n};\n//}}}\n////===\n\n////+++!![PasswordTweak]\n\n//{{{\nversion.extensions.PasswordTweak = {\n major: 1, minor: 0, revision: 3, date: new Date(2006,8,30),\n type: 'tweak',\n source: 'http://tiddlywiki.bidix.info/#PasswordTweak'\n};\n//}}}\n/***\n!!config.macros.option\n***/\n//{{{\nconfig.macros.option.passwordCheckboxLabel = "Save this password on this computer";\nconfig.macros.option.passwordType = "password"; // password | text\n\nconfig.macros.option.onChangeOption = function(e)\n{\n var opt = this.getAttribute("option");\n var elementType,valueField;\n if(opt) {\n switch(opt.substr(0,3)) {\n case "txt":\n elementType = "input";\n valueField = "value";\n break;\n case "pas":\n elementType = "input";\n valueField = "value";\n break;\n case "chk":\n elementType = "input";\n valueField = "checked";\n break;\n }\n config.options[opt] = this[valueField];\n saveOptionCookie(opt);\n var nodes = document.getElementsByTagName(elementType);\n for(var t=0; t<nodes.length; t++) \n {\n var optNode = nodes[t].getAttribute("option");\n if (opt == optNode) \n nodes[t][valueField] = this[valueField];\n }\n }\n return(true);\n};\n\nconfig.macros.option.handler = function(place,macroName,params)\n{\n var opt = params[0];\n if(config.options[opt] === undefined) {\n return;}\n var c;\n switch(opt.substr(0,3)) {\n case "txt":\n c = document.createElement("input");\n c.onkeyup = this.onChangeOption;\n c.setAttribute ("option",opt);\n c.className = "txtOptionInput "+opt;\n place.appendChild(c);\n c.value = config.options[opt];\n break;\n case "pas":\n // input password\n c = document.createElement ("input");\n c.setAttribute("type",config.macros.option.passwordType);\n c.onkeyup = this.onChangeOption;\n c.setAttribute("option",opt);\n c.className = "pasOptionInput "+opt;\n place.appendChild(c);\n c.value = config.options[opt];\n // checkbox link with this password "save this password on this computer"\n c = document.createElement("input");\n c.setAttribute("type","checkbox");\n c.onclick = this.onChangeOption;\n c.setAttribute("option","chk"+opt);\n c.className = "chkOptionInput "+opt;\n place.appendChild(c);\n c.checked = config.options["chk"+opt];\n // text savePasswordCheckboxLabel\n place.appendChild(document.createTextNode(config.macros.option.passwordCheckboxLabel));\n break;\n case "chk":\n c = document.createElement("input");\n c.setAttribute("type","checkbox");\n c.onclick = this.onChangeOption;\n c.setAttribute("option",opt);\n c.className = "chkOptionInput "+opt;\n place.appendChild(c);\n c.checked = config.options[opt];\n break;\n }\n};\n//}}}\n/***\n!! Option cookie stuff\n***/\n//{{{\nwindow.loadOptionsCookie_orig_PasswordTweak = window.loadOptionsCookie;\nwindow.loadOptionsCookie = function()\n{\n var cookies = document.cookie.split(";");\n for(var c=0; c<cookies.length; c++) {\n var p = cookies[c].indexOf("=");\n if(p != -1) {\n var name = cookies[c].substr(0,p).trim();\n var value = cookies[c].substr(p+1).trim();\n switch(name.substr(0,3)) {\n case "txt":\n config.options[name] = unescape(value);\n break;\n case "pas":\n config.options[name] = unescape(value);\n break;\n case "chk":\n config.options[name] = value == "true";\n break;\n }\n }\n }\n};\n\nwindow.saveOptionCookie_orig_PasswordTweak = window.saveOptionCookie;\nwindow.saveOptionCookie = function(name)\n{\n var c = name + "=";\n switch(name.substr(0,3)) {\n case "txt":\n c += escape(config.options[name].toString());\n break;\n case "chk":\n c += config.options[name] ? "true" : "false";\n // is there an option link with this chk ?\n if (config.options[name.substr(3)]) {\n saveOptionCookie(name.substr(3));\n }\n break;\n case "pas":\n if (config.options["chk"+name]) {\n c += escape(config.options[name].toString());\n } else {\n c += "";\n }\n break;\n }\n c += "; expires=Fri, 1 Jan 2038 12:00:00 UTC; path=/";\n document.cookie = c;\n};\n//}}}\n/***\n!! Initializations\n***/\n//{{{\n// define config.options.pasPassword\nif (!config.options.pasPassword) {\n config.options.pasPassword = 'defaultPassword';\n window.saveOptionCookie('pasPassword');\n}\n// since loadCookies is first called befor password definition\n// we need to reload cookies\nwindow.loadOptionsCookie();\n//}}}\n////===\n\n////+++!![config.macros.upload]\n\n//{{{\nconfig.macros.upload = {\n accessKey: "U",\n formName: "UploadPlugin",\n contentType: "text/html;charset=UTF-8",\n defaultStoreScript: "store.php"\n};\n\n// only this two configs need to be translated\nconfig.macros.upload.messages = {\n aboutToUpload: "About to upload TiddlyWiki to %0",\n backupFileStored: "Previous file backuped in %0",\n crossDomain: "Certainly a cross-domain isue: access to an other site isn't allowed",\n errorDownloading: "Error downloading",\n errorUploadingContent: "Error uploading content",\n fileLocked: "Files is locked: You are not allowed to Upload",\n fileNotFound: "file to upload not found",\n fileNotUploaded: "File %0 NOT uploaded",\n mainFileUploaded: "Main TiddlyWiki file uploaded to %0",\n passwordEmpty: "Unable to upload, your password is empty",\n urlParamMissing: "url param missing",\n rssFileNotUploaded: "RssFile %0 NOT uploaded",\n rssFileUploaded: "Rss File uploaded to %0"\n};\n\nconfig.macros.upload.label = {\n promptOption: "Save and Upload this TiddlyWiki with UploadOptions",\n promptParamMacro: "Save and Upload this TiddlyWiki in %0",\n saveLabel: "save to web", \n saveToDisk: "save to disk",\n uploadLabel: "upload" \n};\n\nconfig.macros.upload.handler = function(place,macroName,params){\n // parameters initialization\n var storeUrl = params[0];\n var toFilename = params[1];\n var backupDir = params[2];\n var uploadDir = params[3];\n var username = params[4];\n var password; // for security reason no password as macro parameter\n var label;\n if (document.location.toString().substr(0,4) == "http")\n label = this.label.saveLabel;\n else\n label = this.label.uploadLabel;\n var prompt;\n if (storeUrl) {\n prompt = this.label.promptParamMacro.toString().format([this.toDirUrl(storeUrl, uploadDir, username)]);\n }\n else {\n prompt = this.label.promptOption;\n }\n createTiddlyButton(place, label, prompt, \n function () {\n config.macros.upload.upload(storeUrl, toFilename, uploadDir, backupDir, username, password); \n return false;}, \n null, null, this.accessKey);\n};\nconfig.macros.upload.UploadLog = function() {\n return new config.lib.Log('UploadLog', " !storeUrl | !uploadDir | !toFilename | !backupdir | !origin |" );\n};\nconfig.macros.upload.UploadLog.prototype = config.lib.Log.prototype;\nconfig.macros.upload.UploadLog.prototype.startUpload = function(storeUrl, toFilename, uploadDir, backupDir) {\n var line = " [[" + config.lib.file.basename(storeUrl) + "|" + storeUrl + "]] | ";\n line += uploadDir + " | " + toFilename + " | " + backupDir + " |";\n this.newLine(line);\n};\nconfig.macros.upload.UploadLog.prototype.endUpload = function() {\n this.addToLine(" Ok |");\n};\nconfig.macros.upload.basename = config.lib.file.basename;\nconfig.macros.upload.dirname = config.lib.file.dirname;\nconfig.macros.upload.toRootUrl = function (storeUrl, username)\n{\n return root = (this.dirname(storeUrl)?this.dirname(storeUrl):this.dirname(document.location.toString()));\n}\nconfig.macros.upload.toDirUrl = function (storeUrl, uploadDir, username)\n{\n var root = this.toRootUrl(storeUrl, username);\n if (uploadDir && uploadDir != '.')\n root = root + '/' + uploadDir;\n return root;\n}\nconfig.macros.upload.toFileUrl = function (storeUrl, toFilename, uploadDir, username)\n{\n return this.toDirUrl(storeUrl, uploadDir, username) + '/' + toFilename;\n}\nconfig.macros.upload.upload = function(storeUrl, toFilename, uploadDir, backupDir, username, password)\n{\n // parameters initialization\n storeUrl = (storeUrl ? storeUrl : config.options.txtUploadStoreUrl);\n toFilename = (toFilename ? toFilename : config.options.txtUploadFilename);\n backupDir = (backupDir ? backupDir : config.options.txtUploadBackupDir);\n uploadDir = (uploadDir ? uploadDir : config.options.txtUploadDir);\n username = (username ? username : config.options.txtUploadUserName);\n password = config.options.pasUploadPassword; // for security reason no password as macro parameter\n if (!password || password === '') {\n alert(config.macros.upload.messages.passwordEmpty);\n return;\n }\n if (storeUrl === '') {\n storeUrl = config.macros.upload.defaultStoreScript;\n }\n if (config.lib.file.dirname(storeUrl) === '') {\n storeUrl = config.lib.file.dirname(document.location.toString())+'/'+storeUrl;\n }\n if (toFilename === '') {\n toFilename = config.lib.file.basename(document.location.toString());\n }\n\n clearMessage();\n // only for forcing the message to display\n if (version.major < 2)\n store.notifyAll();\n if (!storeUrl) {\n alert(config.macros.upload.messages.urlParamMissing);\n return;\n }\n // Check that file is not locked\n if (window.BidiX && BidiX.GroupAuthoring && BidiX.GroupAuthoring.lock) {\n if (BidiX.GroupAuthoring.lock.isLocked() && !BidiX.GroupAuthoring.lock.isMyLock()) {\n alert(config.macros.upload.messages.fileLocked);\n return;\n }\n }\n \n var log = new this.UploadLog();\n log.startUpload(storeUrl, toFilename, uploadDir, backupDir);\n if (document.location.toString().substr(0,5) == "file:") {\n saveChanges();\n }\n var toDir = config.macros.upload.toDirUrl(storeUrl, toFilename, uploadDir, username);\n displayMessage(config.macros.upload.messages.aboutToUpload.format([toDir]), toDir);\n this.uploadChanges(storeUrl, toFilename, uploadDir, backupDir, username, password);\n if(config.options.chkGenerateAnRssFeed) {\n //var rssContent = convertUnicodeToUTF8(generateRss());\n var rssContent = generateRss();\n var rssPath = toFilename.substr(0,toFilename.lastIndexOf(".")) + ".xml";\n this.uploadContent(rssContent, storeUrl, rssPath, uploadDir, '', username, password, \n function (responseText) {\n if (responseText.substring(0,1) != '0') {\n displayMessage(config.macros.upload.messages.rssFileNotUploaded.format([rssPath]));\n }\n else {\n var toFileUrl = config.macros.upload.toFileUrl(storeUrl, rssPath, uploadDir, username);\n displayMessage(config.macros.upload.messages.rssFileUploaded.format(\n [toFileUrl]), toFileUrl);\n }\n // for debugging store.php uncomment last line\n //DEBUG alert(responseText);\n });\n }\n return;\n};\n\nconfig.macros.upload.uploadChanges = function(storeUrl, toFilename, uploadDir, backupDir, \n username, password) {\n var original;\n if (document.location.toString().substr(0,4) == "http") {\n original = this.download(storeUrl, toFilename, uploadDir, backupDir, username, password);\n return;\n }\n else {\n // standard way : Local file\n \n original = loadFile(getLocalPath(document.location.toString()));\n if(window.Components) {\n // it's a mozilla browser\n try {\n netscape.security.PrivilegeManager.enablePrivilege("UniversalXPConnect");\n var converter = Components.classes["@mozilla.org/intl/scriptableunicodeconverter"]\n .createInstance(Components.interfaces.nsIScriptableUnicodeConverter);\n converter.charset = "UTF-8";\n original = converter.ConvertToUnicode(original);\n }\n catch(e) {\n }\n }\n }\n //DEBUG alert(original);\n this.uploadChangesFrom(original, storeUrl, toFilename, uploadDir, backupDir, \n username, password);\n};\n\nconfig.macros.upload.uploadChangesFrom = function(original, storeUrl, toFilename, uploadDir, backupDir, \n username, password) {\n var startSaveArea = '<div id="' + 'storeArea">'; // Split up into two so that indexOf() of this source doesn't find it\n var endSaveArea = '</d' + 'iv>';\n // Locate the storeArea div's\n var posOpeningDiv = original.indexOf(startSaveArea);\n var posClosingDiv = original.lastIndexOf(endSaveArea);\n if((posOpeningDiv == -1) || (posClosingDiv == -1))\n {\n alert(config.messages.invalidFileError.format([document.location.toString()]));\n return;\n }\n var revised = original.substr(0,posOpeningDiv + startSaveArea.length) + \n allTiddlersAsHtml() + "\sn\st\st" +\n original.substr(posClosingDiv);\n var newSiteTitle;\n if(version.major < 2){\n newSiteTitle = (getElementText("siteTitle") + " - " + getElementText("siteSubtitle")).htmlEncode();\n } else {\n newSiteTitle = (wikifyPlain ("SiteTitle") + " - " + wikifyPlain ("SiteSubtitle")).htmlEncode();\n }\n\n revised = revised.replaceChunk("<title"+">","</title"+">"," " + newSiteTitle + " ");\n revised = revised.replaceChunk("<!--PRE-HEAD-START--"+">","<!--PRE-HEAD-END--"+">","\sn" + store.getTiddlerText("MarkupPreHead","") + "\sn");\n revised = revised.replaceChunk("<!--POST-HEAD-START--"+">","<!--POST-HEAD-END--"+">","\sn" + store.getTiddlerText("MarkupPostHead","") + "\sn");\n revised = revised.replaceChunk("<!--PRE-BODY-START--"+">","<!--PRE-BODY-END--"+">","\sn" + store.getTiddlerText("MarkupPreBody","") + "\sn");\n revised = revised.replaceChunk("<!--POST-BODY-START--"+">","<!--POST-BODY-END--"+">","\sn" + store.getTiddlerText("MarkupPostBody","") + "\sn");\n\n var response = this.uploadContent(revised, storeUrl, toFilename, uploadDir, backupDir, \n username, password, function (responseText) {\n if (responseText.substring(0,1) != '0') {\n alert(responseText);\n displayMessage(config.macros.upload.messages.fileNotUploaded.format([getLocalPath(document.location.toString())]));\n }\n else {\n if (uploadDir !== '') {\n toFilename = uploadDir + "/" + config.macros.upload.basename(toFilename);\n } else {\n toFilename = config.macros.upload.basename(toFilename);\n }\n var toFileUrl = config.macros.upload.toFileUrl(storeUrl, toFilename, uploadDir, username);\n if (responseText.indexOf("destfile:") > 0) {\n var destfile = responseText.substring(responseText.indexOf("destfile:")+9, \n responseText.indexOf("\sn", responseText.indexOf("destfile:")));\n toFileUrl = config.macros.upload.toRootUrl(storeUrl, username) + '/' + destfile;\n }\n else {\n toFileUrl = config.macros.upload.toFileUrl(storeUrl, toFilename, uploadDir, username);\n }\n displayMessage(config.macros.upload.messages.mainFileUploaded.format(\n [toFileUrl]), toFileUrl);\n if (backupDir && responseText.indexOf("backupfile:") > 0) {\n var backupFile = responseText.substring(responseText.indexOf("backupfile:")+11, \n responseText.indexOf("\sn", responseText.indexOf("backupfile:")));\n toBackupUrl = config.macros.upload.toRootUrl(storeUrl, username) + '/' + backupFile;\n displayMessage(config.macros.upload.messages.backupFileStored.format(\n [toBackupUrl]), toBackupUrl);\n }\n var log = new config.macros.upload.UploadLog();\n log.endUpload();\n store.setDirty(false);\n // erase local lock\n if (window.BidiX && BidiX.GroupAuthoring && BidiX.GroupAuthoring.lock) {\n BidiX.GroupAuthoring.lock.eraseLock();\n // change mtime with new mtime after upload\n var mtime = responseText.substr(responseText.indexOf("mtime:")+6);\n BidiX.GroupAuthoring.lock.mtime = mtime;\n }\n \n \n }\n // for debugging store.php uncomment last line\n //DEBUG alert(responseText);\n }\n );\n};\n\nconfig.macros.upload.uploadContent = function(content, storeUrl, toFilename, uploadDir, backupDir, \n username, password, callbackFn) {\n var boundary = "---------------------------"+"AaB03x"; \n var request;\n try {\n request = new XMLHttpRequest();\n } \n catch (e) { \n request = new ActiveXObject("Msxml2.XMLHTTP"); \n }\n if (window.netscape){\n try {\n if (document.location.toString().substr(0,4) != "http") {\n netscape.security.PrivilegeManager.enablePrivilege('UniversalBrowserRead');}\n }\n catch (e) {}\n } \n //DEBUG alert("user["+config.options.txtUploadUserName+"] password[" + config.options.pasUploadPassword + "]");\n // compose headers data\n var sheader = "";\n sheader += "--" + boundary + "\sr\snContent-disposition: form-data; name=\s"";\n sheader += config.macros.upload.formName +"\s"\sr\sn\sr\sn";\n sheader += "backupDir="+backupDir\n +";user=" + username \n +";password=" + password\n +";uploaddir=" + uploadDir;\n // add lock attributes to sheader\n if (window.BidiX && BidiX.GroupAuthoring && BidiX.GroupAuthoring.lock) {\n var l = BidiX.GroupAuthoring.lock.myLock;\n sheader += ";lockuser=" + l.user\n + ";mtime=" + l.mtime\n + ";locktime=" + l.locktime;\n }\n sheader += ";;\sr\sn"; \n sheader += "\sr\sn" + "--" + boundary + "\sr\sn";\n sheader += "Content-disposition: form-data; name=\s"userfile\s"; filename=\s""+toFilename+"\s"\sr\sn";\n sheader += "Content-Type: " + config.macros.upload.contentType + "\sr\sn";\n sheader += "Content-Length: " + content.length + "\sr\sn\sr\sn";\n // compose trailer data\n var strailer = new String();\n strailer = "\sr\sn--" + boundary + "--\sr\sn";\n //strailer = "--" + boundary + "--\sr\sn";\n var data;\n data = sheader + content + strailer;\n //request.open("POST", storeUrl, true, username, password);\n try {\n request.open("POST", storeUrl, true); \n }\n catch(e) {\n alert(config.macros.upload.messages.crossDomain + "\snError:" +e);\n exit;\n }\n request.onreadystatechange = function () {\n if (request.readyState == 4) {\n if (request.status == 200)\n callbackFn(request.responseText);\n else\n alert(config.macros.upload.messages.errorUploadingContent + "\snStatus: "+request.status.statusText);\n }\n };\n request.setRequestHeader("Content-Length",data.length);\n request.setRequestHeader("Content-Type","multipart/form-data; boundary="+boundary);\n request.send(data); \n};\n\n\nconfig.macros.upload.download = function(uploadUrl, uploadToFilename, uploadDir, uploadBackupDir, \n username, password) {\n var request;\n try {\n request = new XMLHttpRequest();\n } \n catch (e) { \n request = new ActiveXObject("Msxml2.XMLHTTP"); \n }\n try {\n if (uploadUrl.substr(0,4) == "http") {\n netscape.security.PrivilegeManager.enablePrivilege("UniversalBrowserRead");\n }\n else {\n netscape.security.PrivilegeManager.enablePrivilege("UniversalXPConnect");\n }\n } catch (e) { }\n //request.open("GET", document.location.toString(), true, username, password);\n try {\n request.open("GET", document.location.toString(), true);\n }\n catch(e) {\n alert(config.macros.upload.messages.crossDomain + "\snError:" +e);\n exit;\n }\n \n request.onreadystatechange = function () {\n if (request.readyState == 4) {\n if(request.status == 200) {\n config.macros.upload.uploadChangesFrom(request.responseText, uploadUrl, \n uploadToFilename, uploadDir, uploadBackupDir, username, password);\n }\n else\n alert(config.macros.upload.messages.errorDownloading.format(\n [document.location.toString()]) + "\snStatus: "+request.status.statusText);\n }\n };\n request.send(null);\n};\n\n//}}}\n////===\n\n////+++!![Initializations]\n\n//{{{\nconfig.lib.options.init('txtUploadStoreUrl','store.php');\nconfig.lib.options.init('txtUploadFilename','');\nconfig.lib.options.init('txtUploadDir','');\nconfig.lib.options.init('txtUploadBackupDir','');\nconfig.lib.options.init('txtUploadUserName',config.options.txtUserName);\nconfig.lib.options.init('pasUploadPassword','');\nsetStylesheet(\n ".pasOptionInput {width: 11em;}\sn"+\n ".txtOptionInput.txtUploadStoreUrl {width: 25em;}\sn"+\n ".txtOptionInput.txtUploadFilename {width: 25em;}\sn"+\n ".txtOptionInput.txtUploadDir {width: 25em;}\sn"+\n ".txtOptionInput.txtUploadBackupDir {width: 25em;}\sn"+\n "",\n "UploadOptionsStyles");\nif (document.location.toString().substr(0,4) == "http") {\n config.options.chkAutoSave = false; \n saveOptionCookie('chkAutoSave');\n}\nconfig.shadowTiddlers.UploadDoc = "[[Full Documentation|http://tiddlywiki.bidix.info/l#UploadDoc ]]\sn"; \n\n//}}}\n////===\n\n////+++!![Core Hijacking]\n\n//{{{\nconfig.macros.saveChanges.label_orig_UploadPlugin = config.macros.saveChanges.label;\nconfig.macros.saveChanges.label = config.macros.upload.label.saveToDisk;\n\nconfig.macros.saveChanges.handler_orig_UploadPlugin = config.macros.saveChanges.handler;\n\nconfig.macros.saveChanges.handler = function(place)\n{\n if ((!readOnly) && (document.location.toString().substr(0,4) != "http"))\n createTiddlyButton(place,this.label,this.prompt,this.onClick,null,null,this.accessKey);\n};\n\n//}}}\n////===\n\n
__''Chlamy / Flagellar Length Control:''__\n[[axoneme]]\n[[basal body]]\n[[Chlamydomonas]]\n[[IFT]]\n[[rootlets]]\n[[SOFA]]\n[[transition zone]]\n\n__''Microscopy:''__\n[[FIONA]]\n[[FPALM]]\n[[TIRF]]\n[[4Pi]]\n[[Fourier transform]]\n[[Hough transform]]\n[[kymograph]]\n[[speckling]]\n[[cGFP]]\n[[photoactivatable / photoconvertable FPs]]\n
[[Sakowicz98|http://www.sciencemag.org/cgi/content/full/280/5361/292]]
Chlamy mutant with stubby flagella, just past the transition zone. Encodes [[IFT52]] ([[IFT Complex B]])\n[[Brazelton2001|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VRT-448Y7WK-K&_user=4430&_coverDate=10%2F16%2F2001&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=7261f48bd228578c61f4c8487b0ae608]], [[Pete LeFebvre]]\n[[Deane2001|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VRT-448Y7WK-J&_user=4430&_coverDate=10%2F16%2F2001&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=f869ebae94f360563d1a5301a22b018d]], [[Joel Rosenbaum]]
__c__hlamydomonas __G__reen __F__luorescent __P__rotein\n\nDue to difficulties expressing foriegn genes, a ''sythetic GFP'' adapted to [[Chlamydomonas Codon Usage]] was made using oligo ligation. The 30 oligonucleotides used included ''only codons that are most commonly used in well expressed chlamy genes'', and the cryptic intron splice site which leads to misprocessing in several organisms (especially plants) was eliminated. 184 nucleotides out of 714 were changed, which corresponds to a 26% exchange compared to the wild-type gfp Aequorea gene. The ''G + C content'' was thus increased from 39.5% to 61.6%. \n\nIn addition, three amino acid substitutions were introduced: ''S2A'', ''F64L'' and ''S65T''. The latter two mutations made the cGFP comparable to EGFP: 1) when excited at 480 nm the ''fluorescence intensity increased 30-fold''; 2) the chromophore ''formation is accelerated 10- to 20-fold''; 3) the mutant protein shows ''only one excitation maximum at 489 nm'' instead of two (395 and 470 nm).\n[img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_22gnkxghgh]][img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_20djqvr8gv]]\n[[Fuhrmann1999|http://www.blackwell-synergy.com/doi/full/10.1046/j.1365-313X.1999.00526.x?cookieSet=1]]\n\n__''It should be possible to engeneer this into PA-cGFP following the [[Jennifer Lippincott-Schwartz]] example:''__\n\n"To facilitate their use in mammalian cell biology experiments, we generated mutations in a codon-optimized GFP variant developed from the commercially available EGFP. We made __''L64F''__ and __''T65S''__ mutations within EGFP to recover wild-type GFP absorbance properties consisting of a major peak at 397 nm and a minor peak at 475 nm. An additional __''V163A''__ mutation was made to improve folding at 37°C. This construct is referred to as WEGFP. Several T203 mutations, in addition to T203I, were made in this construct which reduced the minor peak and retained the major peak at ~400 nm (Table S1). While many mutants had spectral properties that were comparable with the T203I, the __''T203H''__ mutant was exceptional and decreased the minor absorbance peak even more drastically than the isoleucine mutation. The T203H mutant proved to be an excellent photoactivatable marker and was named [[PA-GFP]]." [[Patterson2002|http://www.sciencemag.org/cgi/content/full/297/5588/1873]]\n\nUse [[Quick Change System|http://www.stratagene.com/manuals/200518.pdf]] to make site-directed changes
stabilizes flagella- prevents readsorption
Severs chlamydomonas flagella at the [[SOFA]], just above the [[transition zone]]\n\nWas used to isolate flagella for the [[chlamy flagellar proteome|http://labs.umassmed.edu/chlamyfp/index.php]].\n[[
[[Merdes2000|http://www.jcb.org/cgi/content/full/149/4/851]]
[[CC-1389 fla1 mt-|http://www.chlamy.org/cgi-bin/ace/generic/tree/default?name=fla1&class=Allele]]\nCannot regenerate flagella at 32 degrees, motility impaired\nLinked to [[fla2]], but is apparently not on [[linkage group IX]]\n[[Mike Adams]] and [[David Luck]]\n\n
Cannot regenerate flagella at 32 degrees, motility impaired.\n[[Mike Adams]] and [[David Luck]]
Cannot regenerate flagella at 32 degrees, motility impaired.\n[[Mike Adams]] and [[David Luck]]
Cannot regenerate flagella at 32 degrees, motility impaired.\n[[Mike Adams]] and [[David Luck]]
Cannot regenerate flagella at 32 degrees, motility impaired.\n[[Mike Adams]] and [[David Luck]]
[[CC-1390 fla2 mt-|http://www.chlamy.org/cgi-bin/ace/generic/tree/default?name=fla2&class=Allele]]\nFlagella are fragile, detach at 32 degrees but can be reassembled. Motility Impaired.\nLinked to [[fla1]], but is apparently not on [[linkage group IX]]\n[[Mike Adams]] and [[David Luck]]
[[CC-1391 fla3 mt-|http://www.chlamy.org/cgi-bin/ace/generic/tree/default?name=fla3&class=Allele]]\nCannot regenerate flagella at 32 degrees, motility impaired\n[[KAP]] non-motor subunit of heterotrimeric kinesin used in antrograde [[IFT]].\n[[linkage group X]] - close to but distinct from [[fla4]]. Single basepair mutation found by [[Mueller05|http://www.molbiolcell.org/cgi/content/abstract/16/3/1341]]\nBackground strain that [[KAP-GFP]] was created in.\n[[Mike Adams]] and [[David Luck]]
Cannot regenerate flagella at 32 degrees, motility impaired.\n[[Mike Adams]] and [[David Luck]]
Cannot regenerate flagella at 32 degrees, motility impaired.\n[[Mike Adams]] and [[David Luck]]
Cannot regenerate flagella at 32 degrees, motility impaired.\n[[Mike Adams]] and [[David Luck]]
Cannot regenerate flagella at 32 degrees, motility impaired.\n[[Mike Adams]] and [[David Luck]]
Cannot regenerate flagella at 32 degrees, motility impaired.\n[[Mike Adams]] and [[David Luck]]
Cannot regenerate flagella at 32 degrees, motility impaired.\n[[Mike Adams]] and [[David Luck]]
[[enh1]]\n[[sun1]]\n[[spf3]]\n[[spf2]]
encodes lf1p, a protein with no homologs that resides within puntate structures in the cell body. [[Nguyen2005|http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1449559]], [[Pete LeFebvre]]
encodes a cytoplasmically localized serine-threonine kinase (STK) [[Pete LeFebvre]]
encodes lf3p, a large (133kD) protein with no sequence homologs that localizes to puntate spots in the cell body. [[Tam2003|http://www.jcb.org/cgi/content/full/163/3/597]], [[Pete LeFebvre]]
encodes a MAP Kinase [[Berman2003|http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VRT-4909176-X&_user=4430&_coverDate=07%2F01%2F2003&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000059594&_version=1&_urlVersion=0&_userid=4430&md5=89c57f10b1aa48119eb1a123aabd533e]], [[Aselon1998|http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=9504917]], [[Pete LeFebvre]]
[[pf1]]
[[spf3]]\n[[mt]]\n[["mut5-47"]]
[[fla3]]\n[[fla4]]
[[Induces flagellar elongation|http://ec.asm.org/cgi/content/full/3/5/1307/F1]].\nFirst described by Nakamura and Kojima, 1987\n[img[alternate text|http://docs.google.com/FilePage?id=dd397pj8_6d2qk82fr]]\n\nLithium treatment depletes the pool of flagellar proteins from the cell body and the [[heterotrimeric kinesin]] Fla10p accumulates in flagella. The kinase activity of [[GSK3]] is inhibited by lithium in vitro. Tyrosine-phosphorylated, active [[GSK3]] is enriched in flagella and GSK3 associates with the axoneme in a phosphorylation-dependent manner. Active GSK3 increases after deflagellation but returns to predeflagellation levels after 30 min.\n[[Wilson2004|http://ec.asm.org/cgi/content/full/3/5/1307]]\n[[Nedra Wilson]], [[Pete LeFebvre]]\n\n20 mM lithium chloride induces ectopic CLASP2 MT lattice binding in the cell body of PtK1 cells (CLASP2 normally only coats MTs in the lamella). Overexpression of GSK3b prevents CLASP2 lattice binding in the lamella. Constituatively active Rac1 induces ectopic CLASP2 MT binding (like lithium), wheras dominant negative Rac1 prevents CLASP2 binding (like GSK3b overexpression)\n[[Wittmann2005|http://www.jcb.org/cgi/content/full/169/6/929]], [[Torsten Wittmann]] and [[Clare Waterman-Storer]]\nInterestingly, CLASPs seem to be important in stabilizing asymmetric MTs originating at the trans-Golgi and going to leading edge. A membrane-bound protein, GCC185, recruits CLASPs to the TGN, where the CLASPs stabilize pre-existing noncentrosomal MT seeds and promote the elongation of MTs from the TGN. CLASP2 preferentially associates with Golgi MTs and CLASP RNAi prevents Golgi MT sormation after [[nocodazole]] treatment.\n[[Efimov2007|http://www.developmentalcell.com/content/article/fulltext?uid=PIIS1534580707001499]]
[[lf1]]\n[[lf2]]\n[[lf3]]\n[[lf4]]
A cell-permeable small molecule inhibitor of the mitotic kinesin, Eg5, arrests cells in mitosis with monoastral spindles. [[Kapoor2000|http://www.jcb.org/cgi/content/abstract/150/5/975]], [[Tim Mitchison]]\n\nThe drug "CK0106023" selectively inhibits Eg5, creating monopolar spindles. [[Sakowicz2004|http://cancerres.aacrjournals.org/cgi/content/full/64/9/3276]]
1) Add .5M acetic acid to drop the pH of the culture to ''pH4.5''\n\n2) Gently mix and Let culture sit one minute.\n\n3) Add .5M KOH to return the pH of the culture to ''pH7.2''\n\nGenerally, to ''1mL'' [[TAP]] (pH7) add ''60uL'' acetic acid and ''65uL'' KOH.\n\nHowever, to ''1mL'' chlamy culture (pH8), add ''60uL'' acetic acid and ''50uL'' KOH.\n\n''***do test run with new media to make sure that these volumes achieve the correct pH***''
[[pf1]]\n[[pf5]]\n[[pf14]]\n[[pf17]]\n[[pf18]]\n[[gli1]]\n[[gli2]]
[[CC-602 pf1 mt-|http://www.chlamy.org/cgi-bin/ace/generic/tree2.pl/default?name=pf1&class=Allele&expand=Strain#Strain]]\n[[CC-1024 pf1 mt+|http://www.chlamy.org/cgi-bin/ace/generic/tree2.pl/default?name=pf1&class=Allele&expand=Strain#Strain]]\n[[CC-260 pf1 thi8 mt+|http://www.chlamy.org/cgi-bin/ace/generic/tree/default?name=CC-260%20pf1%20thi8%20mt%2B&class=Strain]]\n[[CC-1701 ac31 pf1 mt-|http://www.chlamy.org/cgi-bin/ace/generic/tree/default?name=CC-1701%20ac31%20pf1%20mt-&class=Strain]]\n[[CC-2489 pf1 R65|http://www.chlamy.org/cgi-bin/ace/generic/tree/default?name=CC-2489%20pf1%20R65&class=Strain]]\n[[CC-2502 pf1 R84 mt+|http://www.chlamy.org/cgi-bin/ace/generic/tree/default?name=CC-2502%20pf1%20R84%20mt%2B&class=Strain]]\n[[ATCC 30482 C. reinhardtii pf1 mt+|http://www.chlamy.org/cgi-bin/ace/generic/tree2.pl/default?name=pf1&class=Allele&expand=Strain#Strain]]\nThe pf1 mutant lacks spokeheads; five proteins are missing in two-dimensional isoelectric focusing-SDS gels.\n[[linkage group V]], mapped by Ebersold et al. (1962)\n[[David Luck]], [[Elizabeth Harris]], [[Gianni Piperno]]\n\n[[CC-3001 pf1 suppf3 mt-|http://www.chlamy.org/cgi-bin/ace/generic/tree/default?name=sup-pf3&class=Allele]]\nSpoke specific suppressor system, Restores motility to radial spoke-defective mutants without altering spoke defects. [[spf3]]\n[[linkage group VI]]\n[[Huang82|http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=6461414&dopt=Abstract]], [[Bessie Huang]] and [[David Luck]]\n\n
''Photoactivation:'' hitting with certain wavelength light activates fluorescence\n''Photoconversion:'' hitting with certain wavelength light changes fluorescence (ex: green -> red)\n\n[[PA-GFP]] (irreversible photoactivatible GFP)\n\n[[Dronpa]] (reversible photoactivatible green):\n[[KFP]] (reversible photoactivatible)\n\n[[Kaede]] (irreversible photoconvertable green->red) \n[[EosFP]] (irreversible photoconvertable green->red)\n[[KikGR]] (photoconvertable green->red)\n[[PS-CFP]] (photoconvertable cyan->green)
[[Zhu2006|http://pubs.acs.org/cgi-bin/article.cgi/jmcmar/2006/49/i06/html/jm051030l.html]]\n[[Fang1997|http://pubs.acs.org/cgi-bin/article.cgi/bichaw/1997/36/i50/html/bi971363n.html]]\n\nBlocks axonemal (Fang) and cytoplasmic (Zhu) Dynein activities. Binds at an allosteric site and prevents ATP hydrolysis.
[[fla14-1]]\n[[fla14-2]]\n[[pf7]]\n[[pf8]]\n[[shf1-236]]\n[[shf1-253]]\n[[shf1-277]]\n[[shf2-1249]]\n[[shf2-158]]\n[[shf3-1851]]
[[dhc1b-1]]\n[[stf1-1]]\n[[stf1-2]]\n[[bld1]]\n[[bld2]]\n[[bld10]]\n\n
[[fla1]]\n[[fla2]]\n[[fla3]]\n[[fla4]]\n[[fla5]]\n[[fla6]]\n[[fla7]]\n[[fla8]]\n[[fla9]]\n[[fla10]]\n[[fla11]]\n[[fla12]]\n[[fla13]]\n[[fla15]]\n[[fla16]]\n[[fla17-1]]\n[[fla17-2]]\n[[fla18]]\n[[fla19]]\n[[fla20]]\n[[fla21]]\n[[fla22]]\n[[fla24]]\n[[fla27]]\n[[fla28]]\n[[tnr1]]\n[[ts222]]\n[[shf1-253]]\n[["mut5-47"]]
Paralyzed flagella, temperature-sensitive for flagellar assembly. [[pf1]] locus.\nThis mutant makes a flagellar membrane protein that is larger than its wild type counterpart.\n[[Jonathan Jarvik]] and [[Joel Rosenbaum]]
General ATPase inhibitor, extremely toxic. Can inhibit both kinesin and dynein at different concentrations (100uM and 10uM in vitro).\n[img[alternate text|http://upload.wikimedia.org/wikipedia/commons/5/5c/Vanadate_diagram.png]]