We know that everybody has wetlab problems, just like us. And we actually spend most of the summer looking to standardize the Bacillus subtilis transformation protocol. And we did it! So we are presenting you our new standard:

[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Escherichia_coli_MC1061_competent_cells_protocol Escherichia coli MC1061 competent cells protocol]

Two-step Bacillus subtilis transformation procedure

We notice that you not only need a plasmid to transform into B. subtilis, but a plasmid that has form multimers in a RecA+ E. coli and should be flanked by AmyE 5' & 3', so that when the plasmid is transformed into Bacillus, it does not get degradated, and it makes an integration into the genome.