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Long product amplification (1598bp)


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#1 doctorVolos

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Posted 19 May 2009 - 03:10 AM

Hi!!
I'd like to know your opinion about a PCR with a desired product of 1598bp. Should I use elongase or even Taq would be able to do the amplification? What about the PCR reaction as far as it concerns the timing? How long should the extension step last? What about MgCl?
Thank you very much in advance..


#2 tea-test

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Posted 19 May 2009 - 08:13 AM

should be no problem for a normal Taq. Extension time: I would say 1min/kb but just follow the instructions of the Taq manufacturer (also regarding MgCl2).
tea-test: The artist formerly known as Ned Land

#3 doctorVolos

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Posted 20 May 2009 - 01:36 AM

Thanx a million...

#4 mastermi

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Posted 28 May 2009 - 10:29 AM

I would prefer using Pfu-polymerase because of its correction function.
"Standard" Pfu needs about 1 min for 500 bp, whereas Taq only needs 1 min for 1 kb!




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