Exonuclease (3'->5') protected PCR primers

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Skyrre
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Exonuclease (3'->5') protected PCR primers

Post by Skyrre » Fri May 07, 2010 2:11 pm

Hi,

I'm using a DNA polymerase with high 3'->5' exonuclease activity (T4). Now, my problem is that this exonuclease activity is degrading my PCR primers (to make a long story short... and I'm aware that I can't use T4 in the PCR reaction). So I'm wondering if there's a 3'-end modification I can use to make it resistant to degradation, without affecting the extending properties?

I was thinking of using phosphothioate oligos (PTO), but are they totally resistant to degradation? If not, is there some other alternative that is more widely used to prevent this from happening?

Oh - preferably it should be a cheap modification!

Thanks

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JackBean
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Post by JackBean » Fri May 07, 2010 3:18 pm

why can't you use other polymerase?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

Skyrre
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Re:

Post by Skyrre » Mon May 10, 2010 6:43 am

JackBean wrote:why can't you use other polymerase?


The thing is that I need the exonuclease ability as well.

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