How was learning to analysis the results you got back from the PCR and gel electrophoresis, it seem that this could take some practice. As I am writing this question I see the “band intensities were quantified using ImageJ”, so maybe this made your life easier?
Quantifying the results is actually not as hard as it looks! Essentially for our primer validation experiment all we had to verify that the bands are the right size and present. In the gene expression experiment, we had to compare the band brightness.
We would need to get more quantitative results in order to be sure. We could also knock out the gene using CRISPR to determine if DNA damage repair still happens without Rrm2
we compared our data to the Rpp0 gene as there should not be much upregulation of the gene. We also used samples that were never reverse transcribed to make sure there was no contamination
How was learning to analysis the results you got back from the PCR and gel electrophoresis, it seem that this could take some practice. As I am writing this question I see the “band intensities were quantified using ImageJ”, so maybe this made your life easier?
Quantifying the results is actually not as hard as it looks! Essentially for our primer validation experiment all we had to verify that the bands are the right size and present. In the gene expression experiment, we had to compare the band brightness.
How will qPCR and RNAseek contribute to more solidified findings of Rrm2?
Those protocols allow real time quantitative results that require far less interpretation.
What would you need to do to be certain whether or not Rrm2 is involved in DNA damage repair?
We need to be sure that Rrm2 is involved in dna damage repair in order to know whether or not to study it further in the context of cancer
sorry, thought you asked why we should be certain!
What would you need to do to be certain that Rrm2 plays a role in DNA damage repair?
We would need to get more quantitative results in order to be sure. We could also knock out the gene using CRISPR to determine if DNA damage repair still happens without Rrm2
What negative control did you compare your data to?
we compared our data to the Rpp0 gene as there should not be much upregulation of the gene. We also used samples that were never reverse transcribed to make sure there was no contamination