7 thoughts on “C11 – Hunter

    1. We used software programs Primer3plus and BLAST to design forward and reverse primers that would specifically anneal to the Pprs42 gene in the t. thermophila cells genome and then we ordered those primer sequences from a company. We picked the best option from the software by ruling out the ones that had homodimers, differing melting temperatures, or anything that would prevent annealing.

    1. The higher the concentration of agarose used the smaller the pores will be in the gel and therefore the harder it is for larger pieces of DNA to move down the gel. For our primer validation our largest piece of DNA used was 2,000 bp and therefore we decided to use a 1.2% agarose to let our bands be able to run enough for us to visualize them. Because our largest expected DNA length for our second gel (used to our RT-PCR) was 777bp we decided to use a higher concentration of agarose, therefore using a gel with 1.5% agarose. This was to make sure that our bands would not be run off the gel because if that were the case then we wouldn’t be able to visualize the bands.

    1. We would use gene editing mechanisms such as CRISPR/Cas9 to target the Pprs42 gene in the T. thermophila genome and knock it out.

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