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

(video Bacillus protocol)

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.