apricimo Posted February 28, 2011 Posted February 28, 2011 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).
CharonY Posted February 28, 2011 Posted February 28, 2011 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).
apricimo Posted March 1, 2011 Author Posted March 1, 2011 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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