User:Eric Ma/Notebook/iGEM 2010 Lab Prep/2010/05/13: Difference between revisions
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* I will cut Jason's miniprepped DNA because it had the most amount of DNA present. | * 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. | * 4 double digests + 4 single cuts, for further confirmation. | ||
* Volumes used: | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Reagent''' | |||
| align="center" style="background:#f0f0f0;"|'''Control''' | |||
| align="center" style="background:#f0f0f0;"|'''E''' | |||
| align="center" style="background:#f0f0f0;"|'''X''' | |||
| align="center" style="background:#f0f0f0;"|'''S''' | |||
| align="center" style="background:#f0f0f0;"|'''P''' | |||
| align="center" style="background:#f0f0f0;"|'''ES''' | |||
| align="center" style="background:#f0f0f0;"|'''EP''' | |||
| align="center" style="background:#f0f0f0;"|'''XS''' | |||
| align="center" style="background:#f0f0f0;"|'''XP''' | |||
| align="center" style="background:#f0f0f0;"|'''Master Mix''' | |||
|- | |||
| 10X BSA||2||2||2||2||2||2||2||2||2||20 | |||
|- | |||
| 10X Buffer 2||2||2||2||2||2||2||2||2||2||20 | |||
|- | |||
| EcoRI||0||1||0||0||0||1||1||0||0|| | |||
|- | |||
| XbaI||0||0||1||0||0||0||0||1||1|| | |||
|- | |||
| SpeI||0||0||0||1||0||1||0||1||0|| | |||
|- | |||
| PstI||0||0||0||0||1||0||1||0||1|| | |||
|- | |||
| DNA||3||3||3||3||3||3||3||3||3||30 | |||
|- | |||
| sdW||30||30||30||30||30||30||30||30||30||300 | |||
|- | |||
| Total||37||38||38||38||38||39||39||39||39||370 | |||
|} | |||
* 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. | |||
Revision as of 16:43, 13 May 2010
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Restriction Enzyme Test
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