Lab Electrophoesis

Lab Electrophoesis - J. Michael Lindle Gary Odolecki AP...

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J. Michael Lindle Gary Odolecki AP Biology 3/19/10 Introduction But wait a minute…. . What went wrong? What could have gone wrong? There are a number of things that may have skewed the data in this lab. One possibility is that the DNA may have been broken while they were being loaded into the wells. If the DNA strands were broken up into smaller pieces, then they would travel farther down the electrophoresis chamber and throw off the expected results. The concentration of agarose may not have been appropriate for the size of the DNA strands we were working with. A lower concentration of agarose is better suited for sorting larger DNA fragments, while a solution of higher concentration is better suited for sorting smal- ler DNA fragments. (Refer to the picture) Something that may have further thrown off the lab data could have been the voltage that was applied to the electrophoresis chamber. When you are working with DNA fragments that are about 2 kilobases, the best voltage to use is 5 volts for every centimeter that lies in between the electrodes. This could have thrown off the lab data because as the voltage increases, proportionately, the larger DNA frag- ments move a greater distance than the distance travelled by the smaller DNA frag- ments. Something that may have gone wrong has to do with the electrophoresis buffer; although this did not happen in our lab. The electrophoresis buffer does not have much wiggle room in terms of it’s salinity. If the stock’s salinity is too low, the DNA will barely migrate, while if the stock’s salinity is too high, heat will conduct and the agarose gel will melt. The use of contaminated samples would definitely negatively affect the lab results. Two possibilities could have messed up the samples. The first one is a simple technical error
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This note was uploaded on 01/25/2012 for the course BIO 1111 taught by Professor Odolecki during the Fall '10 term at GWU.

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Lab Electrophoesis - J. Michael Lindle Gary Odolecki AP...

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