crude bacterial DNA prep
jxb at bellsouth.net
Sun Aug 14 00:51:23 EST 2005
We do this all the time with E. coli. All you have to do is touch a pipet
tip to the colony and then dip it into your PCR reaction - its all ready to
go. This is our routine method for cloning chromosomal DNA and for verifying
mutants. Incidently, some of the replys state to use a toothpick. Don't use
a wooden tooth pick - use a pipet tip. With toothpicks you will get variable
results as there seem to be Taq inhibitors present in the wood. If you are
not using E. coli you should pick up a colony with a pipet tip and put it
into water and heat as suggested by others.....
"Michael Sullivan" <mlsulliv at wisc.edu> wrote in message
news:mailman.302.1123547527.29381.methods at net.bio.net...
>I want to use PCR to confirm the presence of a large, low copy number
>plasmid from a bacterial strain.
> Can one simply use a small amount of culture directly in a PCR reaction,
> or should one do some sort of DNA prep, even a crude one, prior to PCR?
> If the latter, does anybody have any suggestions for an easy prep to make
> PCR-quality DNA.
> Michael L. Sullivan
> Plant Research Molecular Geneticist
> US Dairy Forage Research Center
> 1925 Linden Drive West
> Madison, WI 53706
> (608) 890-0046 (Phone)
> (608) 890-0076 (FAX)
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