实验动物模型

Harvest cells and count as for passaging. 1x 107 cells in 1 ml are required for each 1 ml cryovial in 10% DMSO.For example:If 1ml cell suspension obtained from harvest contains 50 x 106 cells/ml.:For ...

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 ...

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 ...

Efficient sporulation of BY4743 requires growth as a patch on freshly prepared GNA presporulation plates at 30°C overnight prior to transfer to liquid sporulation medium. (Note: if GNA is inoculated ...

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 ...

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 ...

10X HC (7 Dropout Amino Acid Liquid)2.4 g tyr3.2 g ile2.0 g phe4.0 g glu8.0 g thr4.0 g asp6.0 g val16.0 g ser0.8 g argin final volume of 4L - Autoclave10X YNB58 g YNB (w/o amino acids and w/o ammoni ...

100X YC (7 Dropout Amino Acid Liquid)1.0 g arginine0.5 g aspartic acid0.5 g isoleucine0.5 g phenylalanine0.5 g proline0.5 g serine1.0 g threonine0.5 g valinein final volume of 100 mlstir with low hea ...

COMPLEX MEDIA YPAD medium or Also Called YPD plus AdenineYeast Extract - Peptone - Dextrose plus Adenine medium 6.0 gm yeast extract (Difco) 12.0 gm Peptone (Difco) 12.0 gm Glucose 60 mg adenine hem ...

Yeast Plates Hahn LabLast modified Tue Oct 6 1998Amino Acid Mix (40X) Use 0.6 g/literTyrosine 2 gArginine 4 gSerine 2 gValine 2 gThreonine 4 gIsoleucine 2 gPhenylalanine 2 gAspartic Acid 2 gProline 2 ...

GST Fusion Protein PrepItalics indicate optional steps especially useful for the analysis of untested proteins. GROWTH AND HARVESTING OF BACTERIAAdd 100 ul ampicillin to 100 ml LB. Inoculate and grow ...

GST Fusion Protein Purification from Yeast5 ml overnight culture of your favorite yeast in your favorite medium. Inoculate 50 ml and grow 30o C shaking O/N until OD600 = 0.8 to 1.2. For SCD cultures ...

YEAST TWO-HYBRID SCREEN WITH LIBRARY AND BAIT(The following protocol is for use with the LIBRARY transformation only)Day 1:Grow an overnight culture of a single colony of yeast transformed with bait ...

Table of contents1. Introduction and Background 2. Interaction mating 2.1 Interaction mating - small scale Protocol 1. Mating assay - small scale for tens of different bait or prey proteins. 2.2 Inter ...

Plasmid isolation from yeast (see Clontech YPH p31) Pick colonies into 0.5ml of SD-Leu (or other appropriate SD medium) Vortex for 1min Leave to grow O/N for 18-24h at 30°C 230-250rpm (best in 5ml bi ...

Rather Rapid Genomic PrepHoffman and Winston Gene 19871. Grow 5 ml yeast cultures to saturation.2. Collect cells by centrifugation and resuspend in 0.5 ml of water. Transfer cells to 1.5 ml microfuge ...

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 ...

Beta-gal AssayAdapted from Current Protocols in Molecular Biology1. Grow 5ml YEPD cultures to mid-log phase.2. Centrifuge and resuspend cells in 5ml Z-buffer then place on ice.3. Measure OD600.4. Use ...

Preparation of Yeast DNA Embedded in Agarose PlugsAnja van Brabant(adapted from Iadonato S. P. and A. Gnirke. 1996. RARE-cleavage analysis of YACs. Methods Mol Biol 54: 75-85.) 1. Inoculate a 5 ml cul ...