Yeast Colony PCR

[Arnoud Kal]

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).

Good luck!

Anoud.

Arnoud Kal
Department of Biochemistry
University of Amsterdam
Academic Medical Centre
Meibergdreef 15
1105 AZ Amsterdam
tel. +31-20-5665381/5665382
fax. +31-20-6915519