S-STEM Scholar Rhiannon Giesegh's Blog

 During the extraction, putting the rest of the samples on a gel was not able to see anything. All that was on the gels was a smear. So to see if there was anything that could be done to purify, so we ran a heat gradient of the fragments of right & tet and added left. This was then run from 64-36*C. During this there was also a plasmid extraction done, this gave us a side project confirmation from this plasmid. 

Creating a linear plasmid is a combination of breaking two different plasmids and attaching them together through different processes. One process in an overlap reaction which consists of primers creating a specific site that both fragments have and willmattach to each other. While another will be restriction enzymes which "cut" the plasmid DNA in a specific way to make either clean cuts or sticky ends. These ends are then attached to each other and put through the PCR machine to see if these a work. Presently I work on the overlap reaction, we have had great success with the left and tet fragments as we can recreate the band needed for this project with these fragments multiple times.  There was some inconsistencies with the combination of outright fragment from deinococcus redidurens and the tet fragment from e.coli. The most we can see so far is that the combination of the fragment causes some kind of complex to bunch up and not leave the wells. As such we are testing to s...