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,
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 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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