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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