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Similar to the above electrophoretic mobility shift assay (EMSA) gel KC-PM-ST, our YP-SSB (Yersinia pestis) bound fully by lane 3-4, demonstrating a high affinity for the ssDNA at low concentrations of SSB. It was unfortunate to discover that we would not be able to use the Cy3 fluorophore's excitation/emission wavelengths to determine a precise binding affinity curve because of the gel's artificial enhancement of our intensity measurements. What are some strategies that you would propose for future classes to mitigate this error in the experiment?
Tai Lu
12/7/2015 09:42:54 pm
This particular experiment was interesting. I also had Yersinia Pestis SSB and it was also bound fully in lanes 3 and 4 as well. This meant that there was a high affinity for the ssDNA at low concentrations of SSB as what Mike has explained. Leave a Reply. |
AuthorDr. Edwin Antony and Dr. Sofia Origanti has put together this course. The material presented here are generated by the students of the BIOL 4102 class at Marquette University. (Fall 2015). Improvements can only be made if there is adequate feedback and we thank you in advance for your time. Archives
November 2016
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