direct PCR from bacterial colonies.

Brian_D Lanoil lanoilb at ava.bcc.orst.edu
Wed Aug 9 11:51:48 EST 1995


In article <40852g$n1t at fs2.abdn.ac.uk>, bch127  <bch127> wrote:
>Does anyone have a method for PCr amplification of a plasmid from bacteria
>scraped off a plate into a 96-well plate as an alternative to minipreps, i.e
>how do you make sure that the bacteria lyse ? is 5 minutes at 95 C enough or do
>you have to do anything else ? I'm sure I heard someone talking about this once
>but I cant remember where.
>		Neil Taylor bch127 at abdn.ac.uk
>

I have done PCR directly from colonies in a tube, not on a 96 well 
plate.  The organism I was working with was a gram - bacteria, and there 
was no problem with lysis at 95 C for 5 minutes.  Other organisms with a 
tougher cell wall might be more difficult; I'm not sure.  But I think 
just about any non-thermophilic organism's cells should lyse pretty 
thoroughly at 95 C.  Good Luck!

Brian

*************************************************************************
Brian D. Lanoil   Department of Microbiology
Oregon State University   Nash 222
Corvallis, OR 97331   (503)737-3334
lanoilb at bcc.orst.edu
"Quippy quote here..."
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