Long PCR

[Wayne M. Barnes, Ph.D.]

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