By Andrew G.
Today, we worked mostly on making the recombinant plasmid by adding our CDC14 fungal gene into the PET15B plasmid using Snapgene. We first practiced by inserting the COX1 gene into the PGEX plasmid. Unfortunately, we got very confused about almost everything, such as the 5’ to 3’ direction, which restriction enzymes to use, and especially reverse complements, which are not to be confused with reverse compliments which are what people use to throw shade while pretending to be nice. “Omg that outfit is so vintage!”
One mishap we had was picking the EcoRI restriction enzyme. We noticed halfway through that there is an EcoRI site on our gene so it would get chopped up. Another mishap was we put the reverse complement of our gene into the reverse complement of our plasmid and had two start codons. Luckily, Dr. Jill and the TAs gave us helpful hints when we needed them.
After the ordeal of making our recombinant plasmid, my group unanimously decided to do the primers tomorrow.
Besides the Snapgene shenanigans, we played Scattergory with TA Helen which I won once. We also did a glitchy Labster on bacterial growth curves and my favorite part, a lecture on homology modeling which we will get to do tomorrow!