HELP! PCR using pfu DNA polymerase

Mike Roper roper-m at titania.HSC.Colorado.edu
Fri May 12 19:15:34 EST 1995


In article <johnaris-1005951339550001 at 128.227.67.104>, johnaris at anatomy.med.ufl.edu (John P. Aris, Ph.D.) says:
>
>Dear colleagues,
>
>I have been using the pfu polymerase to amplify a 3-kb fragment, I tried 
>different conditions:
>94 oC 1.5 min
>45, 50, 55 and 60 oC 2 min
>72 oC 6, 7 and 8 min
>10 and 100ng template
>2.5 and 5 units
>for a 50 ul total reaction. Each time I uesd taq as a control. I could obtain
>a 3-kb product from taq but nothing from pfu. If anyone knows how pfu works,
>would you tell me about it? Thanks.
>
>Pei Wu
>Anatomy and Cell Biology
>University of Florida
>(904)392 4563



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