I am trying to investigate what factors can increase the conjugation rate of plasmids in Klebsiella pneumoniae. So far I've tried antibiotics, temperature, oxygen concentrations. I would really appreciate if someone could help me out with my research. Thanks.
You may try conjugation on solid media (such as LB-plates) by streaking the donor and the recipient strain crosswise from liquid cultures and let them grow overnight. Some bacteria transfer better on solid media than in liquid mating mixes. For transconjugants you streak out from the center of the cross where donor and rcipient overlap on your selective or indicator medium.
You may try several timings for the conjugation. In my experience I got transconjugants for some species after 3 hours mating, whereas other gram negatives worked out after overnight conjugation. Selection by using carbon sources is also a good approach, Check out if your acceptor prefers some carbon source, which is not suitable for the donor.
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