Brain Thaw 1

Brain Thaw 1 - Zach Etkind Brain Thaw #1 To identify the...

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Zach Etkind Brain Thaw #1 To identify the gyrase and reverse gyrase proteins you would need to use a Southern Blot and both negatively and positively super coiled circular DNA. The protein samples should be divided into 4 separate samples each. A certain amount of negatively super coiled DNA should be added to 2 test tubes and a certain amount of positively super coiled DNA should be added to the other 2. ATP should be added to one of each set of two test tubes. The test tubes should then be labeled and left to incubate with the ATP for a short period of time. After incubation with ATP, all of the test tubes should be run on an agarose gel. After all the samples have been run the gel should be transferred to a membrane and then incubated with a probe that will label all of the DNA present on the membrane. For the lanes containing positive and negative super coiling, Reverse gyrase, and no ATP, a band should be present towards the end of the gel because super coiled DNA travels faster than relaxed DNA. For the lanes containing DNA gyrase, positive
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