In the Rad51 lanes, we expected to see an increase in expression (a brighter band) in the lanes with DNA damage induced because it is known to be involved in the DNA damage response in Tetrahymena Thermophila. In Ftt18, we expected for there to be bands of the same brightness in both the untreated and treated cells. This is because Ftt18 is known to be expressed in Tetrahymena Thermophila cells and amplify their cDNA and gDNA.
What sorts of results could you expect from a gene knockout that would help provide more conclusive evidence for or against the gene’s involvement in the DNA damage response pathway?
Going into an experiment like this- assuming that Fa2H does play a part in the DNA damage repair pathway, I would expect for there to be issues seen within the DNA damage response of Tetrahymena Thermophila if the gene was knocked out. This is because if a gene were knocked out that is important to the DNA repair process, the response would not work as well and would lead to less DNA repair.
Rad 51 was a positive control in our experiment. This was a positive control in our experiment because we knew that Rad51 was involved in the DNA damage repair pathway in Tetrahymena Thermophila. We knew this because this gene increases expression after DNA damage is induced in this model organism.
Hey Baleigh! Is there any significance to Rad51 working but not Ftt18 if they were both positive controls? I could be missing something. Did you do multiple runs of this gel and get the same result?
There was not a significance to Rad51 working and not Ftt18 besides the fact that it showed that I made an error when pipetting into my tubes for PCR. I most likely included all of the right compounds with the correct amounts of each in the tubes for the Rad51 gene and not for the Ftt18 gene.
Hi Baleigh! Good work. Why do you think that FA2H only showed up in treated cells?
What did you expect to see in the positive control lane on your gel?
In the Rad51 lanes, we expected to see an increase in expression (a brighter band) in the lanes with DNA damage induced because it is known to be involved in the DNA damage response in Tetrahymena Thermophila. In Ftt18, we expected for there to be bands of the same brightness in both the untreated and treated cells. This is because Ftt18 is known to be expressed in Tetrahymena Thermophila cells and amplify their cDNA and gDNA.
What sorts of results could you expect from a gene knockout that would help provide more conclusive evidence for or against the gene’s involvement in the DNA damage response pathway?
Going into an experiment like this- assuming that Fa2H does play a part in the DNA damage repair pathway, I would expect for there to be issues seen within the DNA damage response of Tetrahymena Thermophila if the gene was knocked out. This is because if a gene were knocked out that is important to the DNA repair process, the response would not work as well and would lead to less DNA repair.
Since it was determined that this gene might not play a role in DNA damage response, are you able to determine if this gene has a function elsewhere?
What was the purpose of the Rad51 gene used within the experiment?
Rad 51 was a positive control in our experiment. This was a positive control in our experiment because we knew that Rad51 was involved in the DNA damage repair pathway in Tetrahymena Thermophila. We knew this because this gene increases expression after DNA damage is induced in this model organism.
Hey Baleigh! Is there any significance to Rad51 working but not Ftt18 if they were both positive controls? I could be missing something. Did you do multiple runs of this gel and get the same result?
There was not a significance to Rad51 working and not Ftt18 besides the fact that it showed that I made an error when pipetting into my tubes for PCR. I most likely included all of the right compounds with the correct amounts of each in the tubes for the Rad51 gene and not for the Ftt18 gene.
How did the Ftt18 gene interact with the Fa2H gene, and why did it not show up in the gel?