Which Klenow product to use?! - With or without exo-nuclease activity? (May/03/2006 )

Ok, I just can't get this right... I have cut something with EcoRI, which generates a 5' overhang. I want to blunt this overhang. Which Klenow do I use? DNA Pol 1 Large Klenow fragment or Klenow Fragment (3´→5´ exo–) ?!

Please help! I'm going nuts over this...

Thanks,
Helena

-Helena115-

Anyone? Pleeaaseeee

-Helena115-

I will be using WITH exonuclease for blunting SphI site!!

-Jiang M-

Do you want to use the insert for expression? Select your Klenow based on your plasmid's coding frame (it's got a 4-base overhang, so the frame will shift). If you're using it for something else where the frame doesn't matter, it doesn't matter. Try both and see if either one works better.

-swanny-

Use DNA Polymerase I, Large (Klenow) Fragment. It will solve the problem.

-scolix-

QUOTE (Helena115 @ May 3 2006, 05:29 PM)

Thanks,
Helena

Hi
i think the DNA Pol 1Large Klenow fragment is better! because the Klenow Fragment(3'-5' exo-)may produce a dT salient like Taq .

-sukeagle-