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blunt-end / 3´A overhang cloning


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#1 torte

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Posted 23 April 2010 - 05:31 AM

Hi,
I am going to order a kit for RACE witch contains a cloning kit fro sequencing. I have chosen the GeneRacer from Invitrogen but I don´t know whether to choose the GeneRacer® Kit with SuperScript® III RT and Zero Blunt® TOPO® PCR Cloning kit for Sequencing or the GeneRacer® Kit with SuperScript® III RT and TOPO TA Cloning® kit for Sequencing. Which one would you suggest and why?
Thanks :( T.

Edited by torte, 23 April 2010 - 05:32 AM.


#2 Curtis

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Posted 23 April 2010 - 12:17 PM

Zero Blunt, because according to the manual below the choice of polymerase is up to you. Usually for cloning it is recommended to use Pfu polymerase which has proof reading characteristic and gives less mutation possibility.

http://tools.invitro...neracer_man.pdf

The Taq polymerase in this manual is High Fidelity, still good, but better use Pfu or as stated in the manual Pfx polymerase.




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