miniprep substitute

Kelly C Thome kthome at
Thu Sep 18 07:49:18 EST 1997

Peter Schuchert <schuchert at> 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,

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.


Kelly C. Thome			kthome at
Department of Pathology		phone: 617-732-6581
Brigham and Women's Hospital	fax:   617-732-7449
Boston, MA 02115
Kelly Thome                          <kthome at>  
*Senders of unsolicited email will be charged a $500 proofreading fee.*

More information about the Methods mailing list