miniprep substitute

Kelly C Thome kthome at world.std.com
Thu Sep 18 07:49:18 EST 1997


Peter Schuchert <schuchert at ubaclu.unibas.ch> writes:

>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

In our lab, we screen transformants for inserts by picking up the colony
on a sterile P20 tip and adding it to 10 microliters of sterile PBS, then
adding 1/5 of that to a standard PCR reaction.  We do a 5 minutes at 95
degrees C start, which lyses the cells.  Works every time for us, and is a
*lot* easier than miniprepping all of them and doing digests.  It's pretty
basic, but if you'd like more info, email me.

Regards,
Kelly

Kelly C. Thome			kthome at world.std.com
Department of Pathology		phone: 617-732-6581
Brigham and Women's Hospital	fax:   617-732-7449
Boston, MA 02115
-- 
___________________________________________________________
Kelly Thome                          <kthome at world.std.com>  
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