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Friday, October 24, 2003
Description We all have to shuffle plasmids/constructs in and out of various E. coli strains for amplification and other purposes. This is a simple five (5) minute protocol for the transformation of E. coli cells. (Not recommended for transformation of Ligation reactions!) Procedure If you want to transform E. coli with supercoiled DNA (mini-prep DNA works fine), I highly recommend the following (originally pointed out to me by Mike Benedik, U of Houston): Take an aliquot of your chemically competent strain of choice and thaw on ice (I've used as little as 10 ul of commercially available or homemade competent cells.). Add 0.5 ul (or less) of mini-prep DNA (50 ug/ml is OK) and incubate on ice for 3-4 min. Spread/streak this on a pre-warmed LB plate containing antibiotic (room temp to 37C is fine.). Incubate 18-24 hrs at RT to 37C. That's it. No SOC, no heat shock. Recipes Supplies Competent cells Miniprep plasmid DNA Solid media with antibiotic of your choice Tips (责任编辑：泉水) |
Quickie Transformations
时间:2005-07-18 00:00来源:Scienceboard.net 作者:admin
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