Nde I restriction enzyme

John Brennand john.brennand at gbapr.zeneca.com
Wed Dec 4 07:55:24 EST 1996


martin

we use this enzyme all the time for cutting pcr products at the ATG's to
get them in frame in Coli expression vectors.

Nde can be a problem on the PCR product so we always add R1 or Bam to
the primers thus nnGAATTCATATG....  or nnGGATCCATATG...., cut the PCR
product with R1 or Bam (+ whatever you put at the other end), clone it
into pBluescript (no Nde site) and after finding the correct clone by
sequencing, we lift the Nde (to whatever 3') fragment out into the
expression vector.

Works every time - or your money back

We always use NEBiolabs NdeI and it never fails on the plasmid

john



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