S-Stem Scholar Anthony Luna's Blog

Hello everyone, Beginning with Lab on Tuesday this week, Trevon and I looked to see if our transformation on D. rad was a success but unfortunately, it was not... So we then did a transformation on E. coli to see if it was possibly the plasmid that was not any good but E. coli took it up so that hypothesis is longer considered, for now. Although today, Saturday, I replated the E.coli to see if the plasmid will be retained and the transformation did in fact occur. Next week we will have a final answer.

The transformation Trevon and I carried out last week did not have the results we were hoping for... When we came in we looked at the plates and there was no sign of growth. We believe this is because we placed the cells in the 37-degree Celcius incubator and as a result cooked our cells. The cells should have been placed in the 27-degree Celcius incubator and that is exactly what we did this past week. With that being said, we hope to get some growth on at least one plate when we come back into the lab on Tuesday this coming up week 11-12.

Today, we finished the transformation and now will just be waiting till Monday or Tuesday to see if any colonies have grown and taken the up the plasmid. However, the task we carried out today was actually dispensing the plasmid on to the plates and then placed in the 37C incubator for growth. The two types of plasmids we used were P.rad1 and P.glow. P.rad1 is a plasmid specifically for D.rad and P.glow is plasmid generally used for transformations involving E.coli. We are really hoping D.rad takes up at least one of the plasmids because this is yet to be accomplished in the lab.

In Lab today, Trevon and I actually began the transformation on D.rad. We made twelve plates of TGY(broth) with two types of antibiotics. The first two sets of plates we had dispensed ampicillin in them varying in concentrations to greater increase our chances of the bacteria taking up the plasmid. The last set of plates had the antibiotic chloramphenicol dispensed in them but this concentration was not varied.