实验动物模型

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 ...

Efficient Transformation of Yeast - LiAc MethodDifferent strains transform at different rates with different methods. Some strains transform much better using the electroporation method. Other ones ar ...

Note: Please citeAgatep R. R.D. Kirkpatrick D.L. Parchaliuk R.A. Woods and R.D. Gietz (1998) Transformation of Saccharomyces cerevisiae by the lithium acetate/single-stranded carrier DNA/polyethylene ...

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 ...

Electro-Transformation of YeastDavid AmbergGrow cells to 1X10E8 or OD600 of 1.2-1.3. Spin cells at 5000rpm for 5 min and wash pellets in an equal volume of ice cold water. Wash in 1/2 volume cold wate ...

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 ...

Yeast Genomic DNA IsolationDavid AmbergGrow 100 ml culture to about 1x10 to the 8th Spin 4 x 15 mls of culture down about 3k x 3 min. Resuspend entire pool in about 6 mls ddH2O aliqoute into 1.5ml mic ...

Yeast transformation using lithium acetate (rapid method)Steve HahnLast Modified December 1997This method works well when transforming with a plasmid and is rapid. (for highest efficiency transformati ...

Rhodamine-Phalloidin/Calcofluor StainingDavid AmbergGrow 50 mls of yeast to 5x10E6. Add formadehyde to the media to 4% (33 mls. of 10%). Fix in media at temperature for 10 min. Spin down cells 2-3 K f ...