PCR on whole yeast cells
A.C.Hilton at bham.ac.uk
Wed Jun 8 03:45:34 EST 1994
In article <2sumhu$spf at mercury.dur.ac.uk>, Bert Popping
<Bert.Popping at durham.ac.uk> wrote:
> 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
> 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
I have tried to perform whole cell PCR on isolates of Salmonella and
Campylobacter using 5ul of an overnight culture in Luria broth as the
source of DNA. Even with pre-boiling I still can't seem to get the
Any tips for bacterial wholw cell PCR?
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