PTC Report-Joe Thompson (1).docx - Thompson 1 Joe Thompson Bio Lab 9-11 Thursday Team 4 26 April 2019 Using Functional Genomics to Predict Ability to

PTC Report-Joe Thompson (1).docx - Thompson 1 Joe Thompson...

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Thompson 1 Joe Thompson Bio Lab 9-11 Thursday, Team 4 26 April 2019 Using Functional Genomics to Predict Ability to Perceive Phenylthiocarbamide Introduction A chemist named Arthur L. Fox poured powdered Phenylthiocarbamide (PTC) into a bottle and accidentally spilled some into the room he was in. In this moment, Arthur and his colleague that was also in the room came across a discovery that would later be instrumental in the world of science. Arthur’s colleague tasted the bitterness of the PTC while Arthur tasted nothing at all (Fox, 1932). However, it was not until much later that geneticist were able to connect this to the world of microbiology. Geneticist were able to sequence the genome and provide evidence that there is a specific inherited trait that codes for either the ability or inability to taste the PTC compound. In this experiment, will be tested in a series of both genotypic and phenotypic tests to allow us to make a strong inference on our PTC phenotype and T2R38 genotype and on the link between the two. These connections that will be made will allow for the identification of whether the DNA codes for a “taster” allele, “non-taster” allele, or “weak-taster” allele. Due to the highly accepted idea that the base pairs of the DNA can be used as genes that code for proteins that eventually create the elements of the physical body, there should be an obvious consistency between the genotype and phenotype. Therefore, the results of both the phenotype and the genotype of the PTC trait in any single person's DNA should result in the same phenotypic conclusion (Withers, 2019a). Methods In the first week of this experiment we extracted and isolated our DNA. The cells were extracted by scraping the inside of our cheeks with a toothpick. The toothpick was then twirled in 200 microliters of 5% chelex buffer. 2 microliters of Proteinase K was then added to the mix. The sample was then vortexed, centrifuged at maximum speeds, and boiled. After this was done, we extracted the DNA from the supernatant of our sample with micropipettes and stored it in ice. Next, we added 5 microliters of our DNA to PCR master mix solution that contains water, dNTPs, M gCl 2 , PCR Buffer, Primers, and Taq Polymerase. These primers attached to the bases 547 to 574 and 826 to 850 of our DNA. The DNA was then put in a PCR machine and ran for 3.5 hours in order to amplify that specific region of our DNA (Withers, 2019a). The second week, the DNA was taken out of the ice where it had been since it had been amplified by PCR, and it was added to a microfuge tube that contains the
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Thompson 2 enzyme, Fnu4HI. This is a restriction enzyme that will cut a mutated sequence of bases that comes in the form of “gttgc”. This restriction enzyme action allowed for the resulting DNA after Gel Electrophoresis to be present in only 303 base pairs for a taster allele or
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