The forward and reverse primers we attempted to validate in the first experiment likely did not anneal to HypeC because they were not specific enough to the gene.
Yes! I would still run a preliminary PCR to test primers and an RT-PCR with gel electrophoresis to test gene expression. The only difference would be that cDNA templates previously treated with hydroxyurea would instead be subjected to radiation.
I know that you are assigned the gene to study, but what was the reasoning behind looking into HypeC, specifically? In other words, why is HypeC thought to be involved in the DNA Damage repair pathway?
HypeC is though to be involved in the DNA damage repair pathway because previous research has shown that it is up-regulated during conjugation, a period in which the organism is rapidly cutting up its genome in order to mate. This period requires extensive DNA repair in the organism, so it follows that a gene up-regulated during this time could likely play a role in the repair.
Great question! My best guess is that this research could impact the knowledge surrounding hereditary diseases involving DNA repair such as Bloom’s and Werner’s syndromes.
In testing for other types of DNA damage like radiation would you use the same methods to test for the HypeC gene?
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Why do you think that the primer didn’t ennil to the gel
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The forward and reverse primers we attempted to validate in the first experiment likely did not anneal to HypeC because they were not specific enough to the gene.
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Yes! I would still run a preliminary PCR to test primers and an RT-PCR with gel electrophoresis to test gene expression. The only difference would be that cDNA templates previously treated with hydroxyurea would instead be subjected to radiation.
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I know that you are assigned the gene to study, but what was the reasoning behind looking into HypeC, specifically? In other words, why is HypeC thought to be involved in the DNA Damage repair pathway?
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HypeC is though to be involved in the DNA damage repair pathway because previous research has shown that it is up-regulated during conjugation, a period in which the organism is rapidly cutting up its genome in order to mate. This period requires extensive DNA repair in the organism, so it follows that a gene up-regulated during this time could likely play a role in the repair.
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How long did you run PCR and electrophoresis?
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The thermocycler for PCR was run for about 30-40 minutes for a total of 35 cycles. Gel electrophoresis took about 20-25 minutes.
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What other diseases could your research impact?
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Great question! My best guess is that this research could impact the knowledge surrounding hereditary diseases involving DNA repair such as Bloom’s and Werner’s syndromes.
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Why do you think the Hep C primers didn’t innel to the gene?
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