BAC Vector with SalI in MCS? - (May/05/2006 )
I may be wrong, but from what I can tell the common restriction sites in the multiple cloning site for BAC vectors are EcoRI, BamHI, and HindIII. I am wanting to clone into a BAC vector, however SalI and NotI are the only RE's that are non-cutters in my sequence. Most also use NotI, but only to insert LacZ and the T7 promoter, as well as sequencing primer regions, and these would be gone if I were to clone with NotI. Does anyone know of any BAC vectors that include a SalI site? If not, does anyone have any other suggestions for cloning ~50-kb? Thanks.
Why not introduce SalI and NotI using oligos which include the following sites EcoRI - NotI - SalI - BamHI, using common recombinant DNA techniques ?
(could perhaps be done by PCR also)