I had a lot of fun during my time at the Fred Hutch Explorers Program. The staff was really friendly and helpful and we got to do several cool experiments, with lab work almost every day. Our group practiced pipetting, inoculating agar plates, running gels, and using CRISPR. I met new people and got to work in an actual lab environment, if only for two weeks. The CRISPR experiment was the most enjoyable. It was late in the second week so I got to use everything that I had learned so far in this program and put my skills to the test. First the 1% agarose gel had to be poured, for which we had to prep the gel box to pour the melted agar in. While this substance was cooling we then concocted the mixture of materials that would allow us to run the gel electrophoresis (running electricity through a gel to arrange the substances inside by number of base pairs). We used the Cas9 protein to cut the BRCA1 DNA. The Cas9 used g(guide)RNA implanted in it to then cut the DNA. With five different types of gRNA to use, we each got very separate sequences. Thus each cut at a distinctly different location on the gene and as such would yield two different results within the upcoming gel. This added to the individuality and excitement of the experiment. Next we used the proteinase K enzyme to break down the excess proteins in the mixture. With the BRCA1 gene cut, we then ran a gel with our freshly made agarose and products. When the gel was finished about 35 minutes later it was placed atop a blue light that illuminated it enough for a picture. Through the gel hadn’t turned out quite the way that I had hoped it did show several bands in each lane, supporting my hypothesis that the Cas9 had worked. The DNA had been cut and this showed in the gel as each cut would be made manifest as two seperate bands. The only problem with this experiment was that the bands were too faint, in some places difficult or even impossible to make out with the naked eye. But in hindsight I realized that this problem might be resolved by adding dye, and I decided to do so in future experiments. As one of the final experiments this ushered in the end of my time in the Explorers Program. I am glad for the experience that I gained from these amazing two weeks and I will certainly be applying the principles that I learned here to my future career, Thank You Fred Hutch!
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