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Long PCR (>10kb) polymerase recommendations

Long PCR polymerase

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3 replies to this topic

#1 swanny

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Posted 23 January 2013 - 02:37 PM

Hi all,
One of the researchers here has asked for advice regarding an accurate Pfu pol for long amplicons, typically greater than 10kb. Does anyone have positive suggestions (I don't want this to be a "lousy enzyme that doesn't work" session)?

Cheers,
Swanny
Heart disease kills more women than breast cancer, but heart attack symptoms differ from men's symptoms. Get to know your heart... it could save your life.

#2 phage434

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Posted 23 January 2013 - 03:10 PM

NEB Q5 or Phusion will likely work. I've had good luck in the (distant) past with Roche Extend doing 12-15Kb PCR, but there are likely better alternatives, such as Q5 or Phusion today.

It's critical to design an excellent primer, probably one longer than you would otherwise.

#3 swanny

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Posted 23 January 2013 - 03:24 PM

Thanks for the advice, phage, I'll pass it along.
Love the avatar, by the way!
Heart disease kills more women than breast cancer, but heart attack symptoms differ from men's symptoms. Get to know your heart... it could save your life.

#4 vilperte

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Posted 08 February 2013 - 05:51 AM

I haven't tried to amplify fragments grater than 6kb. But NEB Phusion works very well with my fragments around 6kb. :)

Another one that also works very well is the PfuTurbo Cx Hotstart DNA Polymerase!





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