how to avoid Non-homologous recombination in gene knock-out

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jizhiyuan
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how to avoid Non-homologous recombination in gene knock-out

Post by jizhiyuan » Sun Jun 21, 2009 2:19 am

I'm Knocking out of bacterial genes. The bacterial genome has 7 copies.
I chose a vector with SacB for marker-free knockout. The recombinants are positive for PCR detection. But all of the recombinants are incorrect for southern blotting, the gene still exist in genome. I think non-homologous recombination happened.
Anybody knows what can I do? @[email protected]

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