You mentioned in the Future Directions section that you could test different methods of inducing DNA damage like exposing an organism to radiation. Could you explain how you induced DNA damage in the T. Thermophila in your experiment that led to your conclusion that Piwi2 did not initiate the damage repair process? Thanks!
In this experiment we induced DNA damage using hydroxyurea (HU). Hydroxyurea is a chemical that can induce DNA damage at specific sites of the genome, which allowed us to measure gene expression after inducing damage in this experiment.
We designed primers using the Tetrahymena Genome Database and the Primer3Plus website. We then narrowed down different primer choices based on factors like the length of primers, GC content, and having limited primer secondary structures
I would expect slightly different results. From this experiment, our Rad51 gene didn’t increase in expression after inducing DNA damage as much as we expected it would. This doesn’t let us make a full conclusion about the gene expression change of Piwi2 from this specific experiment. If the experiment was repeated and we saw the expected and larger increase of gene expression with dna damage induced Rad51, then I would say we would see different results with Piwi2 expresssion.
We know that it is involved in DNA damage repair if the gene expression of the specific gene increases in expression after we induce DNA damage. This is because is the presence of DNA damage, this protein will be over-expressed by the cell in order to respond to the damage.
The PCR set up could have had some experimental errors or some reagents, such as the DNA template, could have been denatured, both giving us alternative results.
Great presentation!! I had just one question though: what sources of DNA damage can you induce (other than radiation) to see if Piwi2 can help determine if this gene could be involved in a repair?
curesymposium.org 
You mentioned in the Future Directions section that you could test different methods of inducing DNA damage like exposing an organism to radiation. Could you explain how you induced DNA damage in the T. Thermophila in your experiment that led to your conclusion that Piwi2 did not initiate the damage repair process? Thanks!
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In this experiment we induced DNA damage using hydroxyurea (HU). Hydroxyurea is a chemical that can induce DNA damage at specific sites of the genome, which allowed us to measure gene expression after inducing damage in this experiment.
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How did you design the primers for specifically binding to the Piwi2 gene?
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We designed primers using the Tetrahymena Genome Database and the Primer3Plus website. We then narrowed down different primer choices based on factors like the length of primers, GC content, and having limited primer secondary structures
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Do you think with the limitations using thymol as an antibiotic it would be worth pursuing?
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Sorry don’t respond to this
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Would you expect similar results when you redo the experiment in the future?
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I would expect slightly different results. From this experiment, our Rad51 gene didn’t increase in expression after inducing DNA damage as much as we expected it would. This doesn’t let us make a full conclusion about the gene expression change of Piwi2 from this specific experiment. If the experiment was repeated and we saw the expected and larger increase of gene expression with dna damage induced Rad51, then I would say we would see different results with Piwi2 expresssion.
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How would you know if it is likely involved in DNA damage?
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We know that it is involved in DNA damage repair if the gene expression of the specific gene increases in expression after we induce DNA damage. This is because is the presence of DNA damage, this protein will be over-expressed by the cell in order to respond to the damage.
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What could have been some of the other experimental errors that effected your results?
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The PCR set up could have had some experimental errors or some reagents, such as the DNA template, could have been denatured, both giving us alternative results.
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Great presentation!! I had just one question though: what sources of DNA damage can you induce (other than radiation) to see if Piwi2 can help determine if this gene could be involved in a repair?
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Some other sources of DNA damage we can use are UV radiation and specific alkylating agents.
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