Hi Mr. Burby,

First, I would like to thanks you for this very helpful protocol. I am now doing the Gibson reaction and transformation of MC1061 competent cells. I get colonies after transformation but with no way to check if the plasmid was correctly assembled and if it is in. I tried colony PCR, miniprep followed by PCR or send for sequencing (low yield). None of these worked. Actually, I get a low yield of the plasmid after miniprep. Is that normal? I'm using a miniprep kit from qiagen, do you use a different kit? or trick for the plasmid extraction?
My other question regards the transformation of the Bacillus strains. I'm working with environmental isolates, mainly B. licheniformis. Did you ever tried using this method in other strains other than B. subtilis? Maybe you could give me some feedback.

Many thanks for your time and help,
Kind regards,
Raquel 2/20/2019 2:15:57 AM Reply