Genetic manipulation is easier in some model bacteria such E. coli and B. subtilis. However, we need to study many medical and environment related bacteria isolates. What experiencedo you have to increase the transformation efficiency for non-model bacteria? What are some key things we need to keep in mind?
Daniel, I have a reason to think that some bacteria are very hard-transforming because of their genetic properties, and it is necessary to design special vectors for these strains. Study of native plasmids (if they have any) of such bacteria would help a lot.
I agree with Alex - modifying native plasmids or using special vectors can help your success rates. Electroporation or, if possible, conjugation is typically much more successful than heat-shock transformation. One other often-forgotten consideration is methylation status. Many people will transform with completely demethylated DNA to reduce the risk of host restriction / degradation. However, some bacteria will preferentially restrict fully demethylated foreign DNA as well so it's important to know (if possible) the restriction barrier in your target microbe.
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