During the first months the team worked mostly in the computers with shorts interventions in the lab, mostly for training in the techniques previous, during and after the transformation of bacterial cells.
So we learned first how to propagate bacteria in sterile media, how to prepare media and how to spread cultures in Petri dishes and to save bacteria in frozen vials.
Then we learned how to make competent cells in order to be transformed.
We prepared a series of sterile Petri dishes with Agar – Blood media, then we spreaded cells of the CD41 strain of E. coli just to discover 20 hours later that the bacteria was already mutated, it was lactose negative and we needed for our experiments a lactose positive strain in order to make a correct selection of transformed cells.
A second attempt to reactivate frozen bacteria in culture was done right away, just to discover that the 5 alpha strain was dead and none of the plates developed colonies. Then we raised the concentration of bacterial cells more tan tenfold and we only got results in one of the plates, this second strain had a very low rate of growing, so we had to wait for one more day in the critical last two weeks. Finally, it was detected that this strain was also mutated regarding the lactose genes.
In conclussion we decided not to do any attempt to transform since we will not be able to select accurately the transformed cells; we better acquire virgin strains in our comming trip and start experiments at our return since we need to make the proteins to be expressed by a bacterial cell system in order to comply with our sponsor and the expectations we have raised among the general population. At that time we will only have to start several cultures of the bacteria and then follow the transformation protocol we are showing attached and select transformants in order to collect the proteins by chromatography methods.