Dear friends,
Does anyone knows any Coli Strain available for plasmid with R6Kγ origin (pir gene) and with reduced recombination to increase plasmid stability? I am currently experiencing a very unstable plasmid of R6Kγ origin with lots of repeats containing. A sequence confirmed plasmid transformed back to coli, only a few percent of colonies still keep everything. Most of them lost part of the plasmid. Any information will be helpful.
Thank you.
You need to ensure that you are using recombination-deficient E. coli strain, e.g. with recA1 allele. The most common strains is DH5α λpir. recA is essential for recombination and general DNA repair, hence recA- strain will increase plasmid stability because there will be no recombination between the repeats.
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