What is a possible explanation for UbiconE2 not being involved with DNA damage repair despite ubiquitin conjugation playing a role in DNA damage repair?
Thank you for the amazing question! The specifics are outside of my range of knowledge, but if I had to make an educated guess I would say that UbiconE2 may aid in DNA damage repair passively. It may not be a part of turning on the pathway, but could be involved in degradation within the pathway.
Thank you for the great question! I don’t believe my primers were exactly the cause for decreased expression, however I do believe different primers should be used going forward. My primers create a really large gDNA product and when I designed them I believe I could have found better primers that met the criteria for GC content, that wouldn’t bind elsewhere, and created a smaller gDNA product.
Would you ever consider going to sequence the UbiconE2 gene in its entirety and seeing if you can find any homology to other genes that would give some indication of its function? What else would you explore if you were able to continue your research into this gene?
That’s a wonderful question! UbiconE2 does have homology in other organisms, including humans, and its function is ubiquitin conjugating. If I were to further research into the gene I would definitely look into its role of ubiquitin conjunction to see how to see how critical the gene is to ubiquitin conjugation.
My primers created gDNA that were larger than what we wanted them to be. However, for my research I don’t think they had a large impact. For both my primer validation and gene expression experiments, all of the bands were at the expected lengths. This means that my primers annealed properly and were able to be used for my research. In my reply to Tawanchai Pine I go more in depth on why I would use different primers in the future.
What is a possible explanation for UbiconE2 not being involved with DNA damage repair despite ubiquitin conjugation playing a role in DNA damage repair?
Thank you for the amazing question! The specifics are outside of my range of knowledge, but if I had to make an educated guess I would say that UbiconE2 may aid in DNA damage repair passively. It may not be a part of turning on the pathway, but could be involved in degradation within the pathway.
What primers would you recommend using and why do you think your primers are the reason that UbiconE2 showed no involvement with DNA damage repair?
Thank you for the great question! I don’t believe my primers were exactly the cause for decreased expression, however I do believe different primers should be used going forward. My primers create a really large gDNA product and when I designed them I believe I could have found better primers that met the criteria for GC content, that wouldn’t bind elsewhere, and created a smaller gDNA product.
In future experiments would you use a different primer and why do you believe that there was a decrease in gene expression? Great presentation!
Would you ever consider going to sequence the UbiconE2 gene in its entirety and seeing if you can find any homology to other genes that would give some indication of its function? What else would you explore if you were able to continue your research into this gene?
That’s a wonderful question! UbiconE2 does have homology in other organisms, including humans, and its function is ubiquitin conjugating. If I were to further research into the gene I would definitely look into its role of ubiquitin conjunction to see how to see how critical the gene is to ubiquitin conjugation.
How does the specific primer chosen relate to the research results, was this what you expected to happen?
My primers created gDNA that were larger than what we wanted them to be. However, for my research I don’t think they had a large impact. For both my primer validation and gene expression experiments, all of the bands were at the expected lengths. This means that my primers annealed properly and were able to be used for my research. In my reply to Tawanchai Pine I go more in depth on why I would use different primers in the future.