Protocol for PCR straight from yeast colony?

conibear at molbio.uoregon.edu conibear at molbio.uoregon.edu
Wed Apr 5 12:45:44 EST 1995


John - I tried this protocol for the first time yesterday to prepare yeast
genomic DNA for PCR - it was fast and easy and worked.  Resuspend a yeast
colony (I used a large one) in 60 µL of lysis buffer, incubate at 37C for
60min, then boil for 10min.  Spin for 2 min in microfuge, then use 1µL of
the supernatant in a 10µL PCR reaction. 

Lysis buffer: 1U/µL lyticase, 0.45% (v/v)tween-20, 0.45% (v/v)NP-40, 50mM
KCl, 10mM tris pH 8.3, 1.5mM MgCl, 0.01% (w/v) gelatin.

ref: Kwiatkowski et al (1990) Nucleic Acids Research 18:7191

>In article <susans-010495113900 at triezenberg2.bch.msu.edu>, susans at student.msu.edu (Susan M. Sullivan) wrote:

> I need to verify if the plasmids that I transformed into my S. cerevisae
> are correct.  I know that it is possible to pick a colony of yeast, lyse
> it, and PCR amplify a fragment of DNA using the DNA retrieved from the
> yeast colony as the template.  However, I do not have a protocol for this
> procedure.  If anyone knows a protocol or where to find it please let me
> know. 
> 
> Thanks
> 
> John Stebbins



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