problem cloning a PCR amp. fragment

Nico Dantuma N.P.Dantuma at biol.ruu.nl
Wed Jun 19 07:03:27 EST 1996


Hi  Anwar,

Is it possible that you have an internal restriction site for one of
the two enzymes (do you know the sequence of the amplified fragment)?
If you do have for example a EcoRI site just near your BamHI site you
won't notice any difference in the lenght of your fargment but after
digestion the fargment will contain two EcoRI ends and you surely
won't be able to clone this in your BamHI/EcoRI cut vector.

If you seriously consider this possiblity you can check it by trying
to clone the same fragment in vectors which has been cut with EcoRI or
BamHI.

Nico Dantuma
 

  





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