Unfortunately, as far as I know there is no way to totally guarantee that we get better concentrations. There were a lot of potential areas where the concentrations could have been reduced. This was an issue we were trying to mitigate all semester. Trying the transfection over again was the only method we tried.
What is the benefit of having the cuts ‘off target’ rather than according to a specified cut site in the Golden Gate Method? What are some possible errors that can occur in this process?
The advantage to the off target cut sites allows us to create segments within the plasmids that have overhangs. These overhangs are specific and allow us to insert pieces together like lego pieces.
Unfortunately there wasn’t a whole lot we could to to guarantee that the concentrations of DNA increased. We spent some time this semester trying to figure out why our concentrations were so low. Sometimes the efficacy of the plasmid insertion is just low (maybe because it was so big) and we were not able to get it out.
From my understanding I think it would be a similar process. Once the plasmid is inserted into the E.coli, we then have to use the E.coli to insert it into the soybean plant. There is potential that the insertion into the soybean could be harder, but I am not sure. The process for inserting plasmids into bacteria seems to be more robust, which is why i think it might be a little bit easier.
curesymposium.org 
Super interesting presentation Aidan! What factors would ensure that more concentrations are achieved?
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Unfortunately, as far as I know there is no way to totally guarantee that we get better concentrations. There were a lot of potential areas where the concentrations could have been reduced. This was an issue we were trying to mitigate all semester. Trying the transfection over again was the only method we tried.
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What is the benefit of having the cuts ‘off target’ rather than according to a specified cut site in the Golden Gate Method? What are some possible errors that can occur in this process?
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The advantage to the off target cut sites allows us to create segments within the plasmids that have overhangs. These overhangs are specific and allow us to insert pieces together like lego pieces.
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Why are the off target cuts advantageous in the Golden Gate Method? What are the biggest potential errors with this method?
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Sorry, the site was overloaded with people I think and my original comment wasn’t popping up. Please use the first comment I made.
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Since there was low concentrations of DNA, what would you do differently next time in order to increase it to a normal amount?
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Unfortunately there wasn’t a whole lot we could to to guarantee that the concentrations of DNA increased. We spent some time this semester trying to figure out why our concentrations were so low. Sometimes the efficacy of the plasmid insertion is just low (maybe because it was so big) and we were not able to get it out.
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Since the DNA concentration was low, what would you do differently in the procedure next time in order to increase it to a normal amount?
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Why are the off target cuts advantageous in the Golden Gate Method? What are the biggest potential errors involved with this method?
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Great presentation! How much more difficult is it to insert a vector into Glycine max than E. coli?
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From my understanding I think it would be a similar process. Once the plasmid is inserted into the E.coli, we then have to use the E.coli to insert it into the soybean plant. There is potential that the insertion into the soybean could be harder, but I am not sure. The process for inserting plasmids into bacteria seems to be more robust, which is why i think it might be a little bit easier.
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Great work! I was wondering exactly how collagen would be extracted from Glycine max. Thanks!
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