8 thoughts on “C30 – Mitchell

    1. Hi Delaney,

      Hydroxyurea was used mainly for two reasons. One, it is a very accessible and relatively safe chemical in a laboratory environment, and two, its a chemical that is known to reliably cause DNA damage. It basically provides us with a way to induce controlled DNA damage.

  1. Hi Cosmo! I thought you did a really great job on the entire presentation! I was wondering, you said the bands indicated that the gel electrophoresis was set up correctly. What specifically made this clear, and if the PCR and gel electrophoresis had been set up incorrectly, what would the bands have looked like?

    1. Hi Isabella,

      The different bands on the gels tell us various things about the success of the experiment. For example the loading controls indicate that an equal amount of cDNA was added to each PCR. The negative controls also indicate that there was no contamination during the PCR or gel electrophoresis process, otherwise we would see bands in these places where we did not expect them. Lastly, the presence of the visible bands themselves show us that there were no missing reagents needed to see the results such as SybrSafe.

  2. What were the other controls involved in the experiment and what are you comparing to determine success? Thanks!

    1. Hi Diego,

      In the DNA damage experiment, we had primers amplifying the Rad51 gene in wells that acted as a positive control. These bands showed that DNA damage occurred successfully in the cell, as Rad51 is a gene that is known to increase in expression when DNA damage occurs. Primers amplifying Atpv0c6 were used in wells that acted as a loading control. These bands basically ensure that cDNA was added to each PCR in equal amounts, because the expression of Atpv0c6 remains the same during DNA damage, and therefore we shouldn’t see any change in these bands. Lastly, with each of these sets of wells with each primer set in them, we had two wells with no cDNA in them. These controls were used to ensure that no contamination occurred within the primers, or any of the other reagents used in this experiment.

    1. Hi Alex,

      DNA damage repair applies to the real world, well… because it occurs on its own in the real world. Cells perform DNA damage repair constantly in response to various environmental factors that damage the genome. Studying the genes/proteins involved in these pathways allows us to learn how to manipulate these pathways, which can allow us to manually manipulate these pathways when they are disturbed within cells.

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