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Processive Polymerases

apricimo
in Biochemistry and Molecular Biology

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Hello I am trying to amplify and clone a 10 kb DNA fragment and I am currently using a Pfx platinum kit. I can get fragments <2 kb amplified no problem but I can't get the 10 kb product.

 

Anyone know of a good system to amplify large chunks of DNA? I have a high GC organism I am working with also so I have to use enhancers (FYI I guess).

There are long range (also called extended range) polymerases that you may want to look into (from the usual manufacturers). Note that quite some of them are not proof-reading. But I had used some that were successful for long high GC amplifications. I cannot remember the name off the top of my head, though (it's been a long while).

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There are long range (also called extended range) polymerases that you may want to look into (from the usual manufacturers). Note that quite some of them are not proof-reading. But I had used some that were successful for long high GC amplifications. I cannot remember the name off the top of my head, though (it's been a long while).

 

I'll look into thanks. Sometimes it just takes knowing what to look for (extended range polymerases got me SuperTaq). I will the coolest microbiologist in the lab using SuperTaq :)

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