PCR on whole yeast cells

Bert Popping Bert.Popping at durham.ac.uk
Mon Jun 6 03:25:34 EST 1994


Dear Alvin,

I haven't found any reference in a journal, but there is a summary of this 
topic in the archive. I used this technique last week and I got excellent 
results! The reason why you can't find it in the literature is propbably 
because it's just to simple. 

But here comes the summary:


Let me summarize our current experience:

1.  As many of you have said, THERE IS NO NEED TO ISOLATE DNA.  We have  
found that one toothpick's worth (we use the small end of a disposable  
innoculating loop to avoid contamination), or about 1/2 of a 2 day YPD  
colony, resuspended directly in a 100 microliter PCR reaction, works just  
fine.  There is also no need to preboil; just resuspend and go.  

2.  We have also found that the PCR product is cuttable by at least one  
restriction enzyme (Sal1) DIRECTLY from the tube.  Kind of surprising with  
all those yeast guts swimming around, but there you have it.  Just spin the  
tube to pellet the glop and use the supernatant.

Needless to say, with this kind of ease of use, whole new worlds have been  
opened up to us in terms of genetic screens.  What a great technique.  The  
guy who invented this should get some sort of prize, at least.


--
                                              ... usual disclaimer applies.
________________________________________
 	Dr. B. Popping				                          
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