Transferring pBE110 to S17-1

Last week attempted to transfer the purified pBE110 plasmid into E. coli S17-1 and E. coli DH5a through electroporation. Made electrocompetent E. coli cells (per May 21, 2014 method) and transformed the pBE110 DNA into it. Let incubate at 37C for 1 hour then plated 200 ul onto LBTet15 and incubated overnight.

Results: No colonies from pBE110 formed on DH5a plates. Colonies formed on pUC19 positive control for the S17-1, but not the DH5a. Had some colonies form on S17-1 pBE110 transformants (see picture below).

                                                         
                   Wild type S17-1 on LB only                            S17-1 pBE110 transformants

Colonies do not look like E. coli. Most likely contaminates. Brothed isolated colony of transformants into LBTet15 broth. Incubated at 37C LBTet15 for 48 hours.

After 48 hours nothing grew in the broth. How unfortunate.

Future plans with this approach may need to be re-evaluated.

I haven't been able to update my online lab notebook as much lately as I have been working on a P. larvae project at the ARS USDA building a lot. I will likely not be posting any of the collaboration data that I have been doing over there at the federal building, but will report online about my work with the bacteria.


//EWW