User:Eric Ma/Notebook/iGEM 2010 Lab Prep/2010/05/13

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Restriction Enzyme Test

 * To test restriction enzymes, digest DNA with E+P, X+S, E+S, X+P; in addition to this, do four single cuts.
 * We can tell which enzyme is not working (if it is such) by examining which pairs are not cutting properly.
 * I will cut Jason's miniprepped DNA because it had the most amount of DNA present.
 * 4 double digests + 4 single cuts, for further confirmation.
 * Volumes used:
 * Digest for ~1-2 hrs (not so crucial) at room temp.
 * Note: Initially wanted only 10 µl of water, but in the end, I calculated for 30 µl instead in the master mix for a total of 300 µl of water. Therefore, the reaction may be a bit dilute, which is why I will let this run while I continue writing and rehearsing the podcast instead. The timing shouldn't be too important.
 * Digest ran overnight on ice after ~2h incubation in 37ºC.


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