No bands were present in the case of the negative controls, because there was no DNA template for the primers to anneal to. The lack of bands in figure 3 can be explained by experimental error that occurred while adding the reagents for the RT-PCR.
Knowing more about which genes are involved in the DNA damage response pathway can impact cancer research because when the DNA damage repair pathway does not work, that is when things like cancer are created. Knowing the cause of cancer can help scientists and researchers when trying to find a way to prevent this DNA damage.
The gene specific primers were used in order to detect expression of gene TTHERM_00439320, while the RPPO and Rad51 primers were used as positive controls. The RPPO primers were used because they were intended to anneal to the DNA template, which could show you if you had experimental errors in cases where this control did not work. The Rad51 primers were used to confirm that DNA damage did effectively occur to the Tetrahymena cells, since Rad51 is a gene that is expected to increase in expression following DNA damage.
The experimental error shown in Figure 3 was most likely caused by an error when adding the reagents for the RT-PCR, or in running the gel electrophoresis. We know this because the Rad51 positive control did not show any bands, and the gene specific lanes also did not show any bands despite having bands in the consensus gel.
Why would there be no bands present?
No bands were present in the case of the negative controls, because there was no DNA template for the primers to anneal to. The lack of bands in figure 3 can be explained by experimental error that occurred while adding the reagents for the RT-PCR.
How does knowing these results impact cancer research?
Knowing more about which genes are involved in the DNA damage response pathway can impact cancer research because when the DNA damage repair pathway does not work, that is when things like cancer are created. Knowing the cause of cancer can help scientists and researchers when trying to find a way to prevent this DNA damage.
Why are three different primers used?
The gene specific primers were used in order to detect expression of gene TTHERM_00439320, while the RPPO and Rad51 primers were used as positive controls. The RPPO primers were used because they were intended to anneal to the DNA template, which could show you if you had experimental errors in cases where this control did not work. The Rad51 primers were used to confirm that DNA damage did effectively occur to the Tetrahymena cells, since Rad51 is a gene that is expected to increase in expression following DNA damage.
What most likely caused the experimental error?
The experimental error shown in Figure 3 was most likely caused by an error when adding the reagents for the RT-PCR, or in running the gel electrophoresis. We know this because the Rad51 positive control did not show any bands, and the gene specific lanes also did not show any bands despite having bands in the consensus gel.