For our primer annealing experiment we used water, for the RNA experiment we used both an untreated sample with reverse transcriptase present and a hydroxyurea treated sample without reverse transcriptase present.
In the first case we picked this negative control to try to eliminate the possibility that we had gDNA contamination in places we didn’t want it. In this instance a more complicated negative control was unnecessary.
In the second case, we used the combination of HU-RT and UT+RT to try to ensure that we didn’t have unwanted RNA showing up in our sample lanes. In this instance we used the combination of negative controls specifically to try to make sure we were not contaminating our experimental samples via pipette error and to try to ensure that positive results came from cDNA and not another source, which we would have, hopefully, picked up in one or the other negative control lanes.
What would be your general ideas into the using the mechanism of deoxidized ribose sugars as future genetic research? DNA that involves the use of Uracil?
This is a broader question than I’ve really considered, but generally, I’d say that Rrm2 and its other associated subunits of ribonuclease reductase may have other, undiscovered effects on DNA repair pathways.
This means that they may (or may not) play a regulatory part in potential future gene therapies.
On the cancer front, I’d suggest that it may be beneficial to look at ways to silence Rrm2 since it’s overexpression in various cancers may be related to those cancers ability to rapidly make copies of the diseased cells and repair portions of the genome of those cells. If that’s the case then silencing this gene would potentially slow the progression of the cancer and also possibly make the cancer more susceptible to current anticancer treatments.
The first place that I’d look at is gene therapy in terms of trying to edit the actual genome rather than just adding a plasmid. But it might have some utility in the replication of plasmids in vivo too.
I haven’t seen anything in the literature about that so I honestly don’t know, but since it’s part of the pathway that turns RNA into DNA in the general sense, I would lean/guess towards saying that the answer is “yes” since I haven’t seen anything that suggests it creates a form of DNA that isn’t usable for ssDNA breaks.
That would be yet another avenue for future research, one that I hadn’t considered before you asked this question.
curesymposium.org 
what was your negative control, and why did you use it as opposed to a different negative control?
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For our primer annealing experiment we used water, for the RNA experiment we used both an untreated sample with reverse transcriptase present and a hydroxyurea treated sample without reverse transcriptase present.
In the first case we picked this negative control to try to eliminate the possibility that we had gDNA contamination in places we didn’t want it. In this instance a more complicated negative control was unnecessary.
In the second case, we used the combination of HU-RT and UT+RT to try to ensure that we didn’t have unwanted RNA showing up in our sample lanes. In this instance we used the combination of negative controls specifically to try to make sure we were not contaminating our experimental samples via pipette error and to try to ensure that positive results came from cDNA and not another source, which we would have, hopefully, picked up in one or the other negative control lanes.
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Why was your specific negative control used instead of a different negative control?
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What would be your general ideas into the using the mechanism of deoxidized ribose sugars as future genetic research? DNA that involves the use of Uracil?
LikeLike
This is a broader question than I’ve really considered, but generally, I’d say that Rrm2 and its other associated subunits of ribonuclease reductase may have other, undiscovered effects on DNA repair pathways.
This means that they may (or may not) play a regulatory part in potential future gene therapies.
On the cancer front, I’d suggest that it may be beneficial to look at ways to silence Rrm2 since it’s overexpression in various cancers may be related to those cancers ability to rapidly make copies of the diseased cells and repair portions of the genome of those cells. If that’s the case then silencing this gene would potentially slow the progression of the cancer and also possibly make the cancer more susceptible to current anticancer treatments.
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Besides DNA repair and providing diagnosis and treatment for various cancers, what other field can benefit from your research ?
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The first place that I’d look at is gene therapy in terms of trying to edit the actual genome rather than just adding a plasmid. But it might have some utility in the replication of plasmids in vivo too.
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Great job! Is Rrm2 effective on single stranded DNA breaks as well, or just in double stranded DNA?
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I haven’t seen anything in the literature about that so I honestly don’t know, but since it’s part of the pathway that turns RNA into DNA in the general sense, I would lean/guess towards saying that the answer is “yes” since I haven’t seen anything that suggests it creates a form of DNA that isn’t usable for ssDNA breaks.
That would be yet another avenue for future research, one that I hadn’t considered before you asked this question.
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