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Pulling Out the Mutant Allele

gc genecutl at mendel.berkeley.edu
Wed Feb 23 08:58:43 EST 1994

My lab is trying the old pulling-out-the-mutant-allele trick
for cerevisiae by transforming in a gapped plasmid and looking
for gene replacement in the plasmid.  I've got two questions:
How much flanking DNA homology do you have to have for
efficient gene replacement?
How large can the gap be in the plasmid and still have
this work (we're working with a large gene)?



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