long DNA amplification - (Oct/29/2009 )
Pradeep Iyer on Oct 30 2009, 03:14 PM said:
DRN on Oct 30 2009, 01:55 PM said:
hey no.......increasing the conc. of polymerase is not gonna help!!! going above the optimal level doesn't increase the efficiency of an enzyme.
Ya i am aware of that.. what i meant was if it is not optimum cause then it will get exhausted!!! may be i was not clear
and i still stick to the dNTP's in excess
yeah, u might stick to ur stance on dNTPs in excess, but polymerase might not stick to its template.....point of disagreement, eh????
......lack of clarity this time also, i guess! -DRN-
I have had very good luck with the Roche Expand long range PCR system, and would certainly try it long before doing the work of splicing together many fragments from multiple PCR reactions. PCRs up to 20 Kb are routine.
-phage434-