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will the template of PCR be added A ?


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#1 yen0731

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Posted 19 March 2012 - 01:40 AM

When I use Taq polymerase for PCR, the end of product sequence will be added "A" by Taq ploymerase.

Now I have a question.

My template is about 800 bp. I will want to use PCR product to do TA cloning.

When I use Taq polymerase for PCR, will the template of PCR reaction be added A ?

Because my friend said it that would be added A to the end of template sequence of PCR reaction.

If my friend is right.

I maybe will construct template to TA cloning.

Colud anyone can tell me?

Edited by yen0731, 19 March 2012 - 01:50 AM.


#2 Curtis

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Posted 19 March 2012 - 07:47 AM

when you use Taq polymerase adenine will automatically be added to both 3' ends of your PCR product, no matter what template DNA you use. But the chance of getting A depends on having G and T on the 5' end of your PCR primers too. I can't give you any reference. look up by yourself on Google. However, I never had this problem, and I always ended up with overhang A.

Just don't use Pfu or Pfx polymerase.

#3 yen0731

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Posted 20 March 2012 - 01:55 AM

Thanks Curtis Posted Image




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