error rates in PCR

Michael L. Sullivan mlsulliv at facstaff.wisc.edu
Thu Aug 17 11:04:40 EST 2000


I had read some time ago that for straight Taq, using dNTP's at 50
micormolar rather than 200 micormolar significantly reduced the error rate.
I routinely was doing this, and in general, I really didn't see many (if
any) errors in the PCR products I cloned (although I'll admit this
experience was limited).  I've recently been using a polymerase mix that
has a proofreading enzyme included.  The literature with the product
indicates to use dNTPs at 200 micormolar (as did another brand of a similar
enzyme cocktail).

My question is whether anybody out there has fooled around with these mixes
and tried using less dNTP, and whether that improves the error rate even
more.  I was just wondering if this was something worth trying (in addition
to things like reducing the number of cycles) to get the error rate down
even more.

If no one has any experience doing this, I'll try it next time I do PCR and
see at least if lower dNTP has any effect on product yield.

Thanks,


Mike

Thanks,

Mike

Michael L. Sullivan, Ph.D

U.S. Dairy Forage Research Center
1925 Linden Drive West
Madison WI, 53706

(608) 264-5144 Phone
(608) 264)-5147 Fax


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