I have wild strains of Bacillus, and I'm with real problems in transforming them... I would like to try a protoplast transformation, can anybody suggest a optimized protocol??
A substantial number of wild Bacillus subtilis strains can be transformed by the method of Anagnostopoulos and Spizizen (PMID: 16561900). Most can be transformed with plasmid DNA by the method of Xue et al 1999 J. Microbiol. Meth. 34:183–191. You might also want to consider the method of Meddeb-Mouelhi et al (PMID: 22387342). I have attached links and some PDF files describing my implementation of these methods.
Hello!! I have been working with other stuff lately and there is a long time I'm outta here. I appreciate the answers. I'm really sorry for how long it took me to view them.
Nikunj Daga, I'll send you my email!
Daniel R Zeigler, I've tried some of these with no results, but numbers 2 and 3 I'll try as soon as possible, thanks!
Wiep Klaas Smits, I'm trying to tranform with a linearized integrative DNA, do you have any suggestions?