miniprep substitute

Mark S. Strom strom at research.nwfsc.noaa.gov
Thu Sep 18 16:07:01 EST 1997


Peter Schuchert wrote:

> I heard that colonies of plasmid transformed E. coli
> can be examined for cloned inserts by boiling a little bit
> of the cells and using this for a PCR.
> Could anyone please give me more details on this protocol.
> Thanks in advance,
> Peter

That's basically the protocol.  Toothpick a bit of a colony into 50 ul
of water in a small eppendorf tube, boil 2-3 min, buzz spin for 15
seconds to pellet any debris, and use 5-10 ul in a  standard 25 ul PCR
reaction.

----
Mark S. Strom, Ph.D.                 (206) 860-3377
Utilization Research Div.            (206) 860-3394 (Fax)
Northwest Fisheries Science Ctr.     strom at research.nwfsc.noaa.gov
NMFS/NOAA                            Microbiologist/Volunteer Webmaster
2725 Montlake Blvd. E.               http://www.nwfsc.noaa.gov/
Seattle, WA  98112-2097





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