Yeast colony PCR - who can do it?

doconnor D.I.Oconnor at liverpool.ac.uk
Thu Jun 8 10:13:57 EST 2000


Thomas,

I found that the Yeast Colony PCR protocol from the Cold Spring Harbor Lab 
Course Manual "Methods in Yeast Genetics", 1997 Edition works quite well 
for S.cerevisiae but not so well with all yeast such as Schizosaccharomyces 
pombe. Points to watch out for are the importance of using a very small 
sample of the colony and that the colony is fresh. In addition, it is only
a preliminary result so it is best to confirm the result using a Southern
Blot, etc.

If you cannot get a copy of the method, email me and I will fax you a copy.

Best wishes,

David

Dr David I.O'Connor
Dept of Surgery
Royal Liverpool University Hosptital
UK
Email doconnor at liverpool.ac.uk


In article <8hmndh$jqf$1 at mercury.hgmp.mrc.ac.uk>, leber at protana.com says...
>
>Hi,
>I would like to rapidly screen a large number of yeast colonies form a 
>plate by PCR for the presence of an insert in a plasmid. Working with 
>bacteria, one can simply drop the colony into the PCR mix and off you go. 
>With yeast this does not seem to work or does someone has good experience 
>with this? Alternatively one has to extract the plasmid. Does someone has a 
>very easy method which can be scaled up to do lets say 50 colonies 
>conveniently at the same time? I guess it could also work to only break up 
>the yeast cells and to use the lysate for PCR. Yeast is, however, 
>notoriously difficult to open. Does someone has a quick method?
>Thanks for your help
>Cheers,
>
>Thomas
>
>----------------------------------------------------------------------------
>Dr. Thomas Leber
>Protana A/S
>Staermosegaardsvej 16
>DK-5230 Odense M
>Denmark
>Tel: +45 6315 2059
>Fax: + 45 6315 2040
>

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