Week of 5/28/24-5/30/24 Making Media and Competent Cells


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 the transformed cells. 8 plates of plain R2A were poured and inoculated, 50ul pipetted. 4 plates consisted of only R2A, this was to be our control factor, and 4 plates of R2A with the antibiotic were poured. 2 plates of competent cells were poured, 2 plates without the plasmid were also poured. We made a plethora of plates in order to account for technical error and spares needed in the event something went awry during experimentation. When we return on Monday, we are going to observe our transformed cells of Sonora in the hopes of garnering a successful plasmid transformation. 


Note: The fogginess observed on the microscope lens was due to individuals' cleaning the lens improperly. 



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