S-STEM Scholar Rhiannon Giesegh's Blog

 Found that the heat gradient ran from 54*C to 34*C were the best band through the gel. While there were not as bright on the gel as we would.so there was another gel run. Then the anneling temperature for the PCR reaction was changed to 62*C from now on. Left and tet was put through a heat gradient of 52-70*C. Lead put together the mix of both master mix and of primers in the solution, adding PCR water from there.

 Ran a heat gradient, saw nothing on the gel not even primers.  This canindicate either that something went wrong inthe PCR process or the samples were mixed up when  they went into the PCR reaction. So the other samples were amplified andsent through a gel. Then an isolationof tet(E.coli plasmid, primers C&D) and Left(D.Rad plasmids A&B) fragments. Following this there was a gel extraction. Ran a different heat gradient because the previous one did not produce the correct results. Have tried a few 3 way mixes to see if there will be anyway to attach the three fragments of left tet and right into a three way assemble. So far nothing on any front.