Direct PCR from E. coli colonies (GENOMIC???)
brad at corona.med.utah.edu
Fri Jul 14 13:04:08 EST 1995
In article <1995Jul13.114706.8906 at mbcf>, heath at mbcf.stjude.org
>Has anybody used this successfuly for amplifying a gene from *genomic*
>E. coli DNA? If so, what kind of yield did you get from how many cycles?
>(I'm thinking of trying it, but just want a few pointers first!)
Our lab routinely amplifys genes from E. coli genomic DNA. We have used
pure chromosomal DNA (CsCl pure, Qiagen), PCR kits (Bio-Rad Insta-Prep
matrix), and by the quick resuspend/boil methods that other people have
described in this thread. Generally the success rate is higher with more
pure DNA, probably less salt or other nasties. Cycles and yields are
dependent upon the primers and gene (size, GC content, etc.) amplified, but
generally in my hands 30 cycles gives a decent yield ~ 500 -> 10,000 ng.
Brad Nicholson |"If it worked the first time, it wouldn't be
Department of Pathology | research."...Brad Nicholson
University of Utah | Live from behind the Zion Curtian.
Salt Lake City, UT 84132 |
brad at corona.med.utah.edu |
or: (801)-581-4365 | My opinions are solely my own.
More information about the Methods