primer restriction sites

Andrei Popov ANDREI.POPOV at bbsrc.ac.uk
Wed Aug 30 09:19:59 EST 1995


>Dear Colleagues,
>        I designed a pair of primers which have restriction sites at
>their 5' ends.  I did not add additional bp since I intended to clone
>that into a PCR vector first.  I am wondering if I can directly digest
>them with the enzymes for cloning into an expression vector.  I would
>appreciate your help if you have done that for these two enzymes.
>        The enzymes are: Bam HI and Hind III.
>
>Frank Chen
>
You can: remove primers,
polish the ends (1U T4 pol, 50mkM dNTP 5-10 min at RT),
co-ligate PCR  fragments, then digest with your RE (REs).

regards

Andrei




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