Summer Research Week 1

The first week has been a great learning experience and has allowed me to put my time and focus on my project. I continued to perform PCRs on the DNA strains of Migrant and European strains of S. Paradoxus (my yeast of interest). My advisor, Professor Murphy, ordered new primers that we added into our cleaned PCRs and sent them to get Sanger Sequenced.

The results of the sequences verified the results that appeared when the strains were sequenced using NextGen. After comparing the genome of the PET111 gene, there were three regions in the chromosome of PET111 that had a nucleotide swap between the two populations of yeast. This information supports the hypothesis that there is a genetic difference between the two populations that may have been caused by co-evolution.

One of the next steps for my project includes starting a CRISPR-Cas9. CRISPR is a technique that allows specific genes to be deleted or added into an organism’s chromosome to study the function of the gene. The protocol that we use to perform CRISPER is from a paper called “New Vectors for Simple and Streamlined CRISPR-Cas9 Genome Editing in Saccharomyces cerevisiae” by Marian Laugherty et al.