yeast colony PCR

Hugh Patterton hughg at access.digex.net
Sun Jan 22 13:03:47 EST 1995


>   ROSENKRANTZ at Gems.VCU.EDU writes:
>  Any good protocols? citations?  THanks
>  
>>>>
I have seen a few postings in the past in which a healthy scoop (approx. 4ul) of a *fresh* yeast colony is directly used in a 
standard PCR mix.  I usually incubate approx. 2ul of a fresh yeast colony or 5ul of O/N culture in 100ul TE (pH8.0)/ 1mg/ml 
zymolyase at 30 degrees for 30min, and then use 1ul of the suspension in a standard PCR reaction.  In my experience the 
zymolyase pre-treatment tends to give a 2-5X better signal for equivalent amounts of substrate.

Regards,

Hugh Patterton
hugh at helix.nih.gov
hughg at access.digex.net





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