If I understand correctly, DNA damage is unavoidable and a natural process and it is in a way ‘regulated.’ You used cancer as an example of DNA damage. But, wouldn’t this be an example of ‘unregulated’ DNA damage as the body, at least in my understanding, has no way to counteract these damaged cells? Additionally, would DNA/gene mutations be an example of DNA damage or is this still considered a separate process?
Thank you! We basically wanted to mitigate the possibility of primers annealing to other genes or annealing with each other( forming the dreaded “primer dimer”). I compared these primers to known sequences and selected those with the best chance of working. It turned out that the validated primers we used for this were almost identical to the ones I designed that didn’t work, the forward primer was just 2 base pairs off from mine!
Thanks, Erin! If you look at Figure 1, you’ll see that there are no bands in my gene specific lanes, meaning that the gene wasn’t amplified. IF it HAD worked, you would expect to see 2 bands of similar size next to each other in those lanes.
Since this is the only gene I researched and since gene knockout experiments usually knockout the gene of interest, I would say this one(TTHERM_00242020)
If I understand correctly, DNA damage is unavoidable and a natural process and it is in a way ‘regulated.’ You used cancer as an example of DNA damage. But, wouldn’t this be an example of ‘unregulated’ DNA damage as the body, at least in my understanding, has no way to counteract these damaged cells? Additionally, would DNA/gene mutations be an example of DNA damage or is this still considered a separate process?
Could you further explain why it would be important in conjugation?
Hi Aeryn,
Great Presentation! Can you explain more about how you designed the PCR primers?
Thank you! We basically wanted to mitigate the possibility of primers annealing to other genes or annealing with each other( forming the dreaded “primer dimer”). I compared these primers to known sequences and selected those with the best chance of working. It turned out that the validated primers we used for this were almost identical to the ones I designed that didn’t work, the forward primer was just 2 base pairs off from mine!
Very interesting presentation. Can you explain how you knew the primers did not anneal in the first round of the experiment?
Thanks, Erin! If you look at Figure 1, you’ll see that there are no bands in my gene specific lanes, meaning that the gene wasn’t amplified. IF it HAD worked, you would expect to see 2 bands of similar size next to each other in those lanes.
Can you explain the significance of each labeled lane of the gel electrophoresis?
Do you have any specific genes in mind if you were to move on to gene knockout?
Since this is the only gene I researched and since gene knockout experiments usually knockout the gene of interest, I would say this one(TTHERM_00242020)