Boiling Method for Minipreps

Michael G. Kurilla mgk2r at Virginia.EDU
Fri Jul 30 16:27:03 EST 1993


tencza at med.unc.edu  writes:
> I would like to know if anyone uses the boiling method for plasmid
> minipreps to prepare DNA for sequencing.  I wish to try this method for
> several pBR322 constructs I have recently transformed into HB101.  Any
> hints would be appreciated.
> ^X

We routinely use the boiling lysozyme method.  Prior to
sequencing, we RNase treat the sample and then Strataclean (we
used phenol previoulsy, but the Strataclean resin is just as
good and reduces organic disposal).  The only problem I would
see is your use of pBR322.  My experience is that pBR322
plasmids just give very low yields of plasmid relative to pUC
or pBS.  We have had very good success with cycle sequencing.
If you have that capability I would try that.

Mike Kurilla
University of Virginia
mgk2r at pcmail.virginia.edu



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