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pfu pcr

pfu pcr - Protocols and Methods Forum

pfu pcr - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 08-10-2009, 06:00 PM
Peter Ellis
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Default pfu pcr



If it works with Taq, why do you need to use pfu? What can you do with
a pfu product that you can't do with a Taq product?

Peter
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  #2  
Old 08-10-2009, 08:32 PM
Jayakumar, R
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Default pfu pcr

The existence of 3-5' proof reading exonuclease function and hence lower mispairing might be one reason.
Jay


-----Original Message-----
From: [Only registered users see links. ] [mailto:[Only registered users see links. ].indiana.edu] On Behalf Of Peter Ellis
Sent: Monday, August 10, 2009 2:01 PM
To: [Only registered users see links. ]
Subject: Re: pfu pcr

If it works with Taq, why do you need to use pfu? What can you do with apfu product that you can't do with a Taq product?

Peter
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  #3  
Old 08-10-2009, 09:20 PM
Peter Ellis
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Default pfu pcr

*sigh* Before I get any more replies telling me "might be a fidelity
issue" then yes, I'm aware what pfu is used for. I've also not yet come
across an amplicon, particularly one of only 1.3kb, where you couldn't
just pick half a dozen clones until you get one with the correct sequence.

If there's a particular region that can't amplify properly with Taq (as
Cathal Garvey suggests), then that's something the OP needs to tell us
about so we can make useful suggestions how to optimise for it.

Peter
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