Our lab has optimized a PCR to amplify the envelope gene of HIV-1 cDNA from first round single genome amplification using the Phusion polymerase (Finnzymes). This was a guarenteed PCR that up until recently never gave any problems. We've tried to change the protocol, but nothing has helped. We've tried amplifying old samples that had previously been ampified with the Phusion polymerase, and these too have failed.
Has anyone else experienced problems with this enzyme on routine PCRs?
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Problems with Phusion Polymerase (Finnzymes)
Started by JinalB, Sep 17 2009 06:04 AM
3 replies to this topic
#2
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Posted 17 September 2009 - 06:11 AM
Nope....this enzyme works fine for my applications. But I am up to ordering new enzyme so I am interested in this. Something similar happened here with a cloning kit....we got replacements two times but they changed maufacturer of something in the kit. When they returned to the old one it started working again.
Have you contacted Finnzymes if something is wrong with the lot?
Have you contacted Finnzymes if something is wrong with the lot?
A man cannot be too careful in the choice of his enemies. (Oscar Wilde)
#3
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Posted 17 September 2009 - 06:51 PM
I haven't had probs with Phusion either, and we have just got a new aliquot which is working as normal.
Edited by leelee, 17 September 2009 - 06:52 PM.
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Posted 17 September 2009 - 10:24 PM
gebirgsziege, on Sep 17 2009, 04:11 PM, said:
Nope....this enzyme works fine for my applications. But I am up to ordering new enzyme so I am interested in this. Something similar happened here with a cloning kit....we got replacements two times but they changed maufacturer of something in the kit. When they returned to the old one it started working again.
Have you contacted Finnzymes if something is wrong with the lot?
Have you contacted Finnzymes if something is wrong with the lot?
The new enzymes we've used are Phusion lots 37 and 40. We've also ordered from two different suppliers, and we're waiting for a response from them as to whether they've altered any of the components. So we just thought we'd check on the net to see if anyone else was having similar problems in the interim.
