Direct PCR of Bacterial Colonies

yolanda yangy at rockvax.rockefeller.edu
Sat Aug 12 22:04:14 EST 1995


I use sterilized pipetman tips and haven't had any problems.  It works
really well if I stab a lot of the colony into the PCR mix, not so well if
I stab just a little bit of the colony into the PCR mix.

Steven Poet <poets at ccmail.orst.edu> wrote:
> Be careful, I've read that toothpicks can inhibit the reaction sometimes.
> 
> schoenfeld at MSSCC.MED.UTAH.EDU wrote:
> >
> > Hi -
> > I've seen a few posts on this subject, most of which involve resuspending
> > the colony in a buffer then using part of this suspension as a PCR
> > substrate.  My variation doesn't involve resuspending the colony,
> > minimizing manipulations:
> > 
> > Just stab the colony w/a toothpick and swirl it in your PCR mix.  Do a 3
> > min denaturation step at 94 before cycling.
> > 
> > Robert Schoenfeld
> > Univ. Utah
> > 



More information about the Methods mailing list