crude bacterial DNA prep

JXB jxb at bellsouth.net
Sun Aug 14 00:51:23 EST 2005


Hi Mike,

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.....

Cheers,

JXB




"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.
>
> Thanks,
>
> Mike
> ---
> Michael L. Sullivan
> Plant Research Molecular Geneticist
> US Dairy Forage Research Center
> ARS-USDA
> 1925 Linden Drive West
> Madison, WI 53706
> (608) 890-0046 (Phone)
> (608) 890-0076 (FAX)
> 




More information about the Methods mailing list