User:Wesley J. Houston/Notebook/MAP/2013/03/22: Difference between revisions
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== | ==3/22/13== | ||
* | * Gel purification was performed using the KAH201 plasmids prepped on 2/28, as well as the KAH182 plasmids prepped on 3/21. | ||
** No DNA was present in the second lane, and the concentrations and purities were determined to be too low to complete a ligation. | |||
* Grew more KAH201 liquid cultures. | |||
** Placed colonies in LB broth overnight in shaking 37C incubator. | |||
{| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table --> | |||
|-valign="top" | |||
| <u>Reagent</u> || <u>Volume</u> || | |||
| rowspan="9" | [[Image:Electrophoresis4.jpg |150px|Results of third electrophoresis]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|- | |||
| DNA (plasmid) || 20 μL | |||
|- | |||
| 10x buffer || 3.0 | |||
|- | |||
| enzyme 1 (Pst1) || 1.0 | |||
|- | |||
| enzyme 2 (Xba1 or Spe1)|| 1.0 | |||
|- | |||
| dH<sub>2</sub>O || 5.0 μL | |||
|- | |||
| || 30 μL --> 37°C/ ~15 min. | |||
|} | |||
> Measure conc.'s of gel purified fragments | |||
{| class="wikitable" border="0" cellspacing="3" <!-- [DNA] table --> | |||
|- | |||
| <u>Sample</u> || <u>260/280</u> || <u>ng/μL</u> | |||
|- | |||
| 1. KAH201 (S/P) || 0.32 || 0.851 | |||
|- | |||
| 2. KAH182 (X/P) || 0.385 || 1.064 | |||
|} | |||
Revision as of 22:23, 25 March 2013
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3/22/13
> Measure conc.'s of gel purified fragments
|