Plasmid construction using adaptors.

Christopher W Kafer ckafer at harp.aix.calpoly.edu
Fri Jul 8 16:53:30 EST 1994



We would like to insert a DNA fragment into a vector in which the two have no restriction sites in common i.e. 1 has a BamHI overhang the other a NcoI overhang.

What I would like is a protocol or advice on joining 2 noncompatible cohesive ends using adaptors.  The only protocols I have found are for using adaptors to add restriction sites to blunt end fragments.  

We have the adaptor, two unphosphorylated ssDNA oligos in separate vials.  Where would you suggest I go from here?  Any advice will be greatly appreciated as we are somewhat inexperienced at constructing our own plasmids.  Responding by either email or posting is fine.  Thanks in advance. 



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