flanking 5'-bases on BglI site within a primer

Fernando king-ferdi at gmx.net
Thu Feb 9 12:13:37 EST 2006


according to fermentas there should be no problem if more than two 
additional bases were added...

http://www.fermentas.com/techinfo/re/restrdigpcrii.htm


Josmar wrote:
> Dear all,
> does anyone have experience how many flanking 5'-bases are needed on
> primers that were designed to introduce BglI sites into a pcr product?
> We have good evidence that our pcr-product with 5 additional bases is
> only very poorly cut, even overnight. We kind of rely on a
> quantitatively cut product because it contains a library. The NEB
> catalogue does not give a hint on this. I guess we will try 10
> additional bases next.
> Cheers
> Josmar
> 



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