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PCR + KO TARGETING VECTOR

VINCENT A. FUNARI vfunari at bio.bu.edu
Mon Jan 24 12:59:07 EST 2000


Does anyone know how PCR errors in a targeting vector would effect
recombination frequency.

I have an SV129 mouse Genomic phage clone, that I have been unable to
subclone.

I have experience "PCRing" the targeting region and would like to PCR
and subclone this for the targeting vector.
With pfu or some of the other higher fidelity Taqs, I could achieve an
error rate of less than 1/500 bps.

There might be 20 nucleotide differences in a 10 kb insert.

Any references or experience with using PCR to generate KO targeting
vectors would be greatly appreciated.

Thank you in advance for your time,
Vincent Funari

Ph.D. Candidate in MCBB
Boston University
353-5311, vfunari at bio.bu.edu







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