Yeast Colony PCR
kal at eland.amc.uva.nl
Fri Jan 26 03:02:51 EST 1996
On 25 Jan 1996, Simon Twigger wrote:
> Hi there,
> I am using the Clontech 2 hybrid system and its come time to check my
> postitive colonies. The manual suggests doing a quick/crude plasmid prep
> for each clone, then PCRing off that and then restricting the PCR product
> using AluI and/or HaeIII to try and identify similar clones - thereby
> cutting down on the amount of sequencing to be done.
> I was wondering if there is a protocol for Yeast Colony PCR, like the one
> we regularly use with bacteria, so that I can PCR my product directly
> from a colony without having to go through the plasmid step? I have heard
> that the problem tends to be getting the yeast open and also the genomic
> DNA contamination.
> Has anyone tried this/published anything on this anywhere?
> Thanks for any help anyone can give me
> Simon Twigger,
> Medical College of Wisconsin, Milwaukee.
Dear Simon and netters,
Check out the following reference (which works for me):
A rapid and reliable DNA preparation method for screening a large number
of yeast clones by polymerase chain reaction
Ling et al.,
NAR 23 no 23, p. 4924-4925 (1995).
Department of Biochemistry
University of Amsterdam
Academic Medical Centre
1105 AZ Amsterdam
More information about the Yeast