Is a 5` phosphate needed for homologous recombination?

05 January 2024 1 2K Report

I am a bit confused whether phosphorylated DNA is needed to achieve efficient homologous recombination in e coli. I believe there are methods that use ssDNA oligos or PCR products as inserts for recombineering so these would usually not have a 5` phosphate but there also seems to be some literature that makes it sound like it is beneficial.

Can anyone clarify this, please.

Thanks

Michael J. Benedik

I can not think why a terminal phosphate would matter for homologous recombination. Maybe it helps stabilize the oligo but even for that I can't think of a mechanism.

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