Hey George! qPCR is a more accurate measure of expression because it allows us to see how much starting material is there whereas end point PCR can only measure expression after the last cycle of elongation. qPCR develops a standard curve, which can be used to estimate other concentrations.
Hey Morgan! If the invalidated primers were used for the remaining experiments it is likely that we wouldn’t see any bands at all since the primers wouldn’t anneal to the Twi7 template. qPCR is a more accurate measure of expression because it allows us to see how much starting material is there whereas end point PCR can only measure expression after the last cycle of elongation. qPCR develops a standard curve, which can be used to estimate other concentrations.
How can a treatment be created now knowing that this gene is not highly expressed after induced DNA damage? Does this specific gene play a role in any common diseases / disorders?
Hey Joshua! It would be helpful to have more evidence to confirm for sure that Twi7 isn’t highly expressed. The fact that we didn’t see any bands might just be a sign of the limitations of end-point PCR or it might be the fact that Twi7 isn’t involved in double stranded breaks and could be involved in single stranded breaks. I was more referring to the fact that the overall experiment to seeing if a protein is involved in DNA damage repair would aid in drug development. I didn’t come across any known diseases or disorders that are a result from Twi7 mutations.
curesymposium.org 
How would Quantitative PCR be a better method of analyzing Twi7 expression?
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Hey George! qPCR is a more accurate measure of expression because it allows us to see how much starting material is there whereas end point PCR can only measure expression after the last cycle of elongation. qPCR develops a standard curve, which can be used to estimate other concentrations.
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What results would you get if you used invalidated primers for following experiments? What makes a qPCR a more accurate measure of expression?
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Hey Morgan! If the invalidated primers were used for the remaining experiments it is likely that we wouldn’t see any bands at all since the primers wouldn’t anneal to the Twi7 template. qPCR is a more accurate measure of expression because it allows us to see how much starting material is there whereas end point PCR can only measure expression after the last cycle of elongation. qPCR develops a standard curve, which can be used to estimate other concentrations.
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How can a treatment be created now knowing that this gene is not highly expressed after induced DNA damage? Does this specific gene play a role in any common diseases / disorders?
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Hey Joshua! It would be helpful to have more evidence to confirm for sure that Twi7 isn’t highly expressed. The fact that we didn’t see any bands might just be a sign of the limitations of end-point PCR or it might be the fact that Twi7 isn’t involved in double stranded breaks and could be involved in single stranded breaks. I was more referring to the fact that the overall experiment to seeing if a protein is involved in DNA damage repair would aid in drug development. I didn’t come across any known diseases or disorders that are a result from Twi7 mutations.
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Could there be different kind of DNA damage that might cause the increase expression of Twi7 that wasn’t attempted for the research?
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Hey Ryan! Yes, single stranded breaks might show different Twi7 expression thank double stranded breaks.
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As someone who was not in this lab, was there a specific reason you chose to use the Twi7 gene?
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