实验动物模型

Yeast Colony PCRAkada et al. Biotechniques 28:668-674 (April 2000)MATERIALS0.25% SDS10X Colony PCR Buffer: 0.125 M Tris-HCl pH 8.50.5625 M KCl25 mM MgCl210 mM dNTP's20% Triton X-100Taq polymeraseTwo G ...

Gene Disruption via PCR 14:115-132 (1998).Reaction mix:5 µl 10X Taq Buffer5 µl 25 mM MgCl22 µl 10 mM dNTP's10-100 ng template DNA25 pmols of each primer0.5 µl Taq polymerase (2 Units)_________________ ...

Quick Yeast DNA Prep: Isolation of Total DNA (genomic and plasmid) Linda Hoskins Hahn Lab 10/16/98Grow a 5 ml YPD O/N culture inoculated with a single yeast colony at 30 deg. Transfer culture to a sma ...

Genomic DNA Miniprepgrow 5 ml cells over night at 30 °C spin wash once with 1 ml H2O resuspend in 500 µl lysis buffer add acid washed glass beads to 1.25 ml vortex 2 min recover liquid phase with blue ...

Yeast Genomic DNA Prep Linda Hoskins/Hahn lab Aug 18 1997(modified from Philippsen 1991)Grow 10ml YPD cultures o/n. Figure out cell density; inoculate 30 ml YPD and grow o/n so that cell density is ap ...

Pick a medium size yeast colony (~3 mm dia) and transfer to 200 ul of lysis buffer. Add an equal volume of glass beads (0.45 mm dia). Mix on vortex at top speed for 1 min. Add 200 ul of phenol/CHCl3 ...

Yeast Miniprepfrom Robzyk and Kassir Nucleic Acid Res. (1992) 20:3790Materials:• 10 ml overnight culture yeast in drop out media (must be very dense)• STET (8% sucrose 50 mM Tris pH 8 50 mM EDTA 5% Tr ...

High Efficency Yeast Plasmid Rescue(by Trey Powers Fields Lab) Grow yeast overnight.Spin down 1.5 ml of culture in microfuge tubes.DecantAdd 250 µl of Qiagen buffer P1 to each tubeAdd about 250 µl of ...

Yeast Prep for FACS 7(13):4335-4346; 1988.1. Spin down 1E7 cells in a microfuge tube for 1 minute.2. Resuspend pellet in 1 ml of 70% EtOH. Fix for at least 60 minutes at room temperature (or up to sev ...

Yeast Cell LysatesModified from the Rine LabGrow cells to 0.3 to 0.5 O.D.(600) (~1E7 cells/ml). Use about 1.0 to 5.0 OD of cells per lysate; if your cells are at 0.5 OD per ml use 2 ml to give you 1.0 ...

Preparation of Yeast Protein Extracts ...

Yeast Protein Isolation "Yaffe-Schatz"David AmbergAdapted from Yaffe M.P. and Schatz G. PNAS 1984 81 4819-4823.Grow 25mls yeast cells to 5x10E6. Sequentially spin down cells in a 15 ml polypro tube. W ...

Yeast Nuclear Extract (Small Scale) Hahn Lab (adapted from J. Leatherwood Ptashne lab)last modified Mon Aug 27 2001Cell Growth3 liters of cells are grown in YPD (3% glucose) to A600 of 3-5. Antifoam ...

Yeast RNA Miniprepgrow cells to mid-log phase (10-20 ml) (or take samples from time courses 5-25 ml) spin cells discard supernatant wash cells once with 1 ml cold RNA buffer freeze pellet in dry ice ( ...

Pol III whole cell yeast extract (small scale)Mike Schultz Pavel Aprikian Steve Hahnlast modified6/28/99Grow 2 liters yeast in YPD (with 3% glucose) to an A600 of 1.5 to 2.0 (inoculate ~1.5 ml satura ...

Curing strains of endogenous 2-micron plasmidIn order to create a circle-zero derivative from a Cir+ strain the method of Rose and Broach (1990) is used. A 2-micron-based plasmid YEp351-GAL-FLP1 * tha ...

Rapid Yeast Protein Prep for SDS PAGE and WesternHorvath and Riezman Yeast 1994; Gottschling LabSample Buffer: 10 ml0.06M Tris-HCl pH 6.80.6 ml 1M Tris 6.810% (v/v) glycerol2 ml 50% glycerol2% (w/v ...

EMS Mutagenesis of Yeast Linda Hoskins Hahn Lab Last modified10/16/981. Grow a 50 ml O/N YPD yeast culture at 30 degrees C.2. Determine cell density and transfer an amount of culture corresponding to ...

ß - GAL Filter Assaystreak yeasts to single colony replica plate onto NC-filter (leave 10 sec.) use tweezers to lift filter and slowly submerge in liquid Nitrogen for 1 min place on back side of a pet ...

Making library DNA from the DNA we send youThe original library was distributed as 18 individual pools. After 8/98 there is a new library that is distributed as 10 pools. You will be sent about a mi ...