实验动物技术

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 ...

Sporulation and Dissection of Yeast a/alpha Diploids Linda HoskinsLast Modified Tue Nov 6 2001These conditions work well for the Hieter yeast strains1. Isolate diploid colonies. Inoculate 2 ml YPD + A ...

Alternate Sporulation Method for BY4743 Protocol: 1. Patch cells that have been freshly grown on YPD to freshly prepared GNA pre-sporulation plates for 1 day at 30°C.Repatch to another freshly prepar ...

Yeast Smash & Grab DNA Miniprep based on M.D. Rose F. Winston and P. Hieter (1990) Methods in Yeast Genetics: A Laboratory Course Manual. Cold Spring Harbor Laboratory Press Cold Spring Harbor New Yor ...

Phenol/Freeze RNA PrepSchmitt et al. (1990) NAR 18 3091-3092.DEPC is used to rid solutions of RNases. DEPC is very toxic and should be used only in the hood.Stuff you need:(DEPC treatment means: add D ...

Yeast Chromosomal DNA Prepuse 25 ml cells per timepoint / sample add cells to 25 ml 100 % Ethanol (at -20 °C) add 1 ml 0.5 M EDTA store at -20 °C thaw cells wash once with H2O resuspend in 0.5 ml Shpe ...

Materials required:Please Note: You Do Not have to Sonicate the Carrier DNA ANYMORE! WHY? I have done the experiments and shown that that Bigger is Better for Carrier DNA! Follow instruction below.Si ...

High efficiency LiAc transformationModified from Gietz Homepage.Stuff you need:10X LiAc 1 M LiAc filter sterilize50% PEG 3350 filter sterilize1. Inoculate 2-5 mls of liquid YPED or 10 ml synthetic med ...

I. SUMMARY The yeast two-hybrid system provides a relatively straight forward approach to understanding protein function. Section II outlines the basic components of ...

This Protocol allows You to prepare Frozen Yeast cells that are competent for transformation after thawingAfter Dohmen et al. (1991) Yeast 7: 691-692. See Schiestl et al (1993) The ability to make yea ...

Large-scale yeast transformationThe following protocols were developed by Petra Ross-Macdonald for Mike Snyder's LacZ fusion project at Yale University. Yeast Cell PreparationThis protocol merges a ty ...

Multiwell Transformation ProtocolLabor-saving implementsMultichannel pipettors--1 to 25 µl and 20 to 200 µl volume (Brinkmann 12-channel). Sterile 96-well assay or tissue culture plate with lid (such ...

Typical transformation resultsThe following transformations were performed at the same time using the multiwell transformation protocol. Similar amounts of PCR product were used for each. The pictures ...