Ligation question

William O'Connell woconnel at nmh.org
Mon Feb 1 12:44:01 EST 1999


I just had another idea:  Can I use an internal 5' biotinylated primer and
unlabeled 5 and 3' primers in the pcr reaction?  Can I purify the pcr
product with an internal biotin label?  I think this way I will have more
problems in the pcr step and none in the ligation step, whereas my original
idea had no problems in the pcr step and more problems in the ligation
step.  Any other ideas?
Thanks again. BIll

William O'Connell <woconnel at nmh.org> wrote in article
<01be4e06$ffb0cde0$dd3814a5 at WOCONNEL.NMH.ORG>...
> I have a biotinylated pcr product which I can purify using streptavidin
> magnetic beads.  I want to clone the product by ligating it into a T/A
> vector.  Will the beads or the biotin interfere with the cloning step? 
> Should I simply use unlabeled primers in my pcr reaction?
> Thanks in advance.  Bill
> 



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