实验动物模型

Beta-galactosidase Reporter Gene Assay (Liquid Form) REFERENCE: Hoffman G. Garrison T. R. and Dohlman H. G. Analysis of RGS proteins in Saccharomyces cerevisiae Methods Enzymol. 344:617-631 2002. ...

mTn-3xHA/GFP TRTn3 terminal inverted repeatsXaFactor Xa cleavage recognition siteloxRlox site target for Cre recombinaseGFPgene encoding Green Fluorescent Protein mutant p11URA3URA3 gene from S. cerev ...

PLATE Transformation"PLATE" is an acronym of the names of ingredients in the transformation solution: PEG Lithium Acetate Tris and EDTA.-Use sterile technique and sterile solutions throughout this met ...

Direct PCR from Whole Yeast Cells: Zymolyase MethodContributor: Namjin ChungDate: June 18 19961. An average-size yeast colony (0.5-2mm) or a cell pellet from a liquid culture is touched with asterile ...

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 ...

This Protocol allows for TRAFO with any Yeast cell sourceReferenece; Gietz R.D. and R.A. Woods. (2002) TRANSFORMATION OF YEAST BY THE Liac/SS CARRIER DNA/PEG METHOD. Methods in Enzymology 350: 87-96.W ...

SMALL Scale Transformation 1. Inoculate 1 ml of YPDA or SD with several 2�3 mm colonies. 2. Vortex vigorously to disperse any clumps. 3. Transfer cells to a flask containing 50 ml of YPDA or SD: 4. I ...

HIGH EFFICIENCY TRANSFORMATIONReference: Gietz R.D. and R.A. Woods. (2002) TRANSFORMATION OF YEAST BY THE Liac/SS CARRIER DNA/PEG METHOD. Methods in Enzymology 350: 87-96.All Solutions for the LiAc/SS ...

Yeast Transformationinoculate 5-10 ml culture with single colony and grow ON measure OD600nm and dilute to OD600nm = 0.3 in 50 ml grow for 2-4 hrs spin wash cells with 25 ml H2O spin resuspend in 2 ml ...

LiOAc YEAST TRANSFORMATIONa(see notes at end)Use STERILE TECHNIQUE throughout.1. Inoculate a 10 ml overnight culture with a single yeast colony. If starting with a yeast strain with no plasmids use YP ...

Yeast Transformation (high efficiency) Linda Hoskins/Hahn LabLast modified Wed Jul 5 2000This more involved method gives higher transformation efficiency than the semi rapid method. Use DMSO if transf ...

Yeast Transformation Using Frozen Competent Cells and Single-stranded DNA as a CarrierKatherine KolorReferences: Schiestl and Gietz (1989) Curr. Genetics 16:339-346 Gietz et al (1995) Yeast 11:355-360 ...

DNA isolation from yeast 1) Start a 5 ml o/n culture 2) Pellet o/n culture in a purple screw cap tube (this may take several spins) and dump s/n.3) Add 100 µl of STET to pellet and vortex briefly.4) A ...

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 ...

Joel Huberman's DNA isolation procedure with modifications by Bonny Brewer1. Kill cells at mid log phase by adding Na Azide to 0.1%. Immediately chill culture by adding it to frozen 0.2 M Na EDTA. (4 ...

Sucrose Gradient Protocolfrom Song et al. 1996 Modified by Ginger HoffmanGrow a 2 ml culture 24 hr. at 30oC in selective media When culture is ready use it to inoculate about 55 ml (50 ml plus 5 for ...

Whole Cell ExtractsREFERENCE: Hoffman G. Garrison T. R. and Dohlman H. G. Analysis of RGS proteins in Saccharomyces cerevisiae Methods Enzymol.344:617-631 2002. -Use sterile technique and sterile so ...

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 ( ...

Yeast mini whole cell extract using glass beads for western analysisSteve HahnLast modified Fri Oct 16 1998Before trying this method try rapid protein prep which involves boiling yeast in SDS PAGE buf ...

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 ...