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Removal of 5 overhangs to form blunt ends


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#1 maringa

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Posted 09 June 2010 - 11:47 AM

Hi all,

Am having problems with a new construct and I need to put my protein of interest in frame, I need to remove the 5' overhangs generated when I digest with XhoI. Unfortunately, T4 polymerase exonuclease activity is 3'-5', I know Polymerase I has 5'-3' exonuclease activity but am confused, I don't know how to use it to cut these 5' overhangs, please HELP :wacko:

Thanks
John

#2 pDNA

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Posted 09 June 2010 - 12:01 PM

DNA polymerase I can not be used to remove 5'-overhangs ...use Mung bean nuclease instead!

Here you can find how to do it.

Regards,
p




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