Wayne M. Barnes, Ph.D.
barnes at BIODEC.WUSTL.EDU
Wed Mar 16 14:12:02 EST 1994
Dear Genetic Technologists,
Update to my fresh PNAS paper (91:2216-2210) on Long and Accurate PCR.
For best results with 28-35 kb, be careful not to overdo the cycling.
My most recent conditions are .1 or 1 ng of lambda DNA per 100 ul (incubate
as 33-50 ul), and only 16 cycles. 1.2 ul KlentaqLA enzyme mix.
If you cycle too much, the product band goes to heck, which is a place in
the origin well of the gel from which it can never return.
For 8 kb or so, use about half as much enzyme, 0.6 ul. -- Wayne
Barnes at biodec.wustl.edu -------------------------
Wayne M. Barnes, Ph.D. fax: 314/362-7183
Dept. Biochemistry 8231, Washington Univ. Med.School
St. Louis, Missouri 63110 USA ph: 314/362-3351
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