实验动物模型

Preparation of single-stranded probes from cloned cDNA(from Patel and Goodman 1992)a) 2-5 µg of plasmid DNA containing the cDNA insert is linearized using an appropriate restriction enzyme. For antise ...

MMS integration (both methods modified by Herman Lab) NOTES: Remember that MMS is a potent mutagen -- wear gloves and be very careful. Integration is best done using an array that is transmitted at ve ...

Nematode dye filling(Ed Hedgecock ) Thumb plate Transfer worms into a known amount of M9 in a labeled depression. Add DiO(2 mg/ml in DMF) at 10 l/ml of M9. Let sit for 2 hours. Transfer worms with a P ...

Whole-mount in situ hybridization for the detection of RNA in C. elegans embryosGeraldine Seydoux and Andrew Fire Adapted from Seydoux G. and Fire A. (1995). Whole-mount in situ hybridization for the ...

"CONVENTIONAL" TWO STEP FIXATION 1995 1. Wash animals in M9 and anaesthetized in 8% alcohol in M9 5’ 2. 2.5% glutaraldehyde 1% paraformaldehyde in 0.1M sucrose 0.05M cacodylate on ice 2 hours 3. 3 ri ...

Phalloidin Staining worms by Michael Koelle modified from Beth Bucher and Andrew Chisholm 4/6/94 Phalloidin binds to filamentous actin. This stain allows visualization of mainly of muscles but some ot ...

Staining C. elegans for ß-galactosidase activity using X-gal by Michael Koelle adapted from Laird Bloom Jeff Way & Michael Basson 4/6/94 1. Grow up a small plate of worms. Some protocols call for usin ...

Embryo BuffersEgg Salts118 mM NaCl 48 mM KCl 2 mM MgCl2 2 mM CaCl2 10 mM HEPES (pH 7.4) ...

Cuticle preparationsThis procedure was adapted from one described over the phone (thanks P.G.). We're not sure if it appears elsewhere in published form.-Collect embryos on apple juice-agar plates for ...

GENERAL MAINTENANCE INSTRUCTIONS FOR IMAGINAL DISC CELL LINES This is our lab protocol for growing imaginal disc cells (clone-8) The Milner Lab website has additional protocols for clone-8 cells. Cl ...

Medium Shields and Sang Medium 3 (SS3) modified for low serum. Supplemented before use with 2% heat inactivated FCS 2.5% FE2 12.5IU/100ml insulin and filter-sterilised through 0.22µm filter. Supplemen ...

SINGLE-FLY DNA PREPS FOR PCRGloor et al. 1993 Genetics 135:81-95We have developed a simple method for the rapid and reproducible isolation of DNA from single flies for amplification by the polymerase ...

IMMUNOHISTOCHEMISTRY ON Drosophila BRAINS Caution! In converting these protocols to HTML some units have been scrambled.Specifically microliters (ul) often appear as milliliters (ml). Think first! 1. ...

Labelling Drosophila pupae with BrdU ITO Kei 1992 OctoberCOLLECTING STAGED PUPAE 1. Prepare a couple of bottles containing late 3rd instar larvae which wander on the botlle wall (110-120 hours after e ...

Non-fluorescent protein/RNA double-labeling(a version of this is described in: Manoukian A. and Krause H.M.(1992) Genes Dev. 6: 1740-1751.)1. Collect embryos dechorionate and fix in a (1:1) mixture o ...

Single and Double FISH protocols for DrosophilaThis procedure is taken from a recently published book chapter. The reference is: Hughes S. and Krause H.M. (1998) Single and double FISH protocols for D ...

Yeast ProtocolsRather Rapid Genomic Prep Plasmid Rescue Protein Prep Yeast Histone Prep (from Jakob Waterborg; .pdf format) Media RecipesHC mediaHC Dropout Plate Supplementation Table YC mediaYC Dropo ...

June 1997 - optimized by N. Kertesz L.Leyns & E. De RobertisPlease cite: Belo J.A Bouwmeester T. Leyns L.Kertesz N. Gallo M. Follettie M. and De Robertis E.M. "Cerberus-like is a secreted factor with ...

Freezedown Guidelines What to SaveFor each 96-pack each lab will have yeast representatives from all four strains: BY4741 (or BY4730) BY4742 (or BY4739) BY4743 and the homozygous diploid. It is recogn ...

FEDEX Protocol(Or: How to increase the likelihood that your FEDEX packages reach their intended destination in a timely fashion)While this list seems pedantic so far it only incorporates actual bad ex ...