S-STEM Scholar Madi Cook's Blog

Returning to lab on Monday, we found that another lab mate's competent cells we had utilized for our project was contaminated so we had to scrap all of our bacterial specimens that was used with those cells. This set us back quite a bit, however we were able to bounce back. In the future, we intend on using our own supply of any cells we may need in order to reduce the risk of contamination again due to technical errors. It was established that we will be making R2A media every Monday, preferably 250ml of such in order to comfortably have a steady line of media. On Tuesday, we took the OD value of Sonora using R2A as our blank, with which we received an OD value of 1.63. On Wednesday we inoculated 600 micro liters of our bacterial culture with colony PCR in order to verify that our transformation of Sonora was successful. E. Coli was our positive control, while Sonora would be our negative control. This is because E. Coli contains the plasmid PRAD1 already, which is what we are try...

We are currently working with Deinococcus Sonora in the hopes of transforming the bacteria with PRAD1 plasmid, PRAD1 being the general plasmid that is utilized the most due to it's ability to be accepted easier compared to that of other plasmids. Returning to lab on Tuesday, we curated 250ml of R2B media and 250ml of R2A media. Within the R2A media there were two additional flasks, one containing the antibiotic Chloramphenicol with a concentration of 3mg/ml. The other flask contains R2A media without any antibiotic present. On Wednesday we made competent cells and inoculated those in order to engage in the transformation attempt of Sonora. We also poured 20 plates, and made more R2A media (250ml) to autoclave and be ready for use when we return to lab on Monday. Gram stains were performed on the transformed cells of Sonora to ensure no contamination was present, with which we garnered gram positive results, shape being that of purple spherical-short rods, that confirmed our use of ...