Difference between revisions of "Team:NYU Shanghai/Protocols"
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− | + | Link to <a href="https://www.neb.com/tools-and-resources/usage-guidelines/optimizing-restriction-endonuclease-reactions"> NEB Protocol </a> | |
+ | |||
+ | <br><br>To determine buffer for double digests, we used: <a href="https://www.neb.com/tools-and-resources/interactive-tools/double-digest-finder">NEB Protocl</a> | ||
+ | |||
+ | <br><br>Guide to heat inactivation: <a href="https://www.neb.com/tools-and-resources/usage-guidelines/heat-inactivation">NEB Protocol</a> | ||
+ | |||
+ | <br><br>General reaction set-up: | ||
+ | <table border="1"> | ||
+ | <tr> | ||
+ | <td>Restriction enzyme</td> | ||
+ | <td>10units, generally 1uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DNA</td> | ||
+ | <td>1 ug</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>10X NEB Buffer</td> | ||
+ | <td>5uL (1X)</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Total Reaction Volume</td> | ||
+ | <td>50uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Incubation Time</td> | ||
+ | <td>1hr</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Incubation Temperature</td> | ||
+ | <td>Enzyme Dependent</td> | ||
+ | </tr> | ||
+ | </table> | ||
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Revision as of 14:07, 16 September 2015