HELP! PCR using pfu

[John P. Aris, Ph.D.]

Dear colleagues,
I have been using pfu DNA polymerase to amplify a 3-kb fragment from yeast 
genomic DNA. I tried different conditions:
94 oC 1.5 min
45,50,55,60 oC 2 min
72 oC 6,7,8 min
10 and 100ng template
2.5 and 5 units 
for a total of 50ul reaction. Each time I used taq polymerase as a control.
I could obtain a 3-kb product from taq but nothing from pfu. If anyone knows
how pfu works, would you please tell me about it? Thanks.

Pei Wu
Anatomy and Cell Biology
University of Florida
(904)392-4563