PCR on whole yeast cells
Bert.Popping at durham.ac.uk
Mon Jun 6 03:25:34 EST 1994
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
University of Durham
Dept. Biol. Sci. PHONE +44 91 374 2430
South Road FAX +44 91 374 2417
Durham DH1 3LE E-MAIL Bert.Popping at durham.ac.uk
WWW set pointer to: http://www.dur.ac.uk/~dbl1bp
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