User:Wesley J. Houston/Notebook/MAP/2013/03/25: Difference between revisions
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== | ==3/25/13== | ||
* | * Prepped plasmids of KAH201 and determined concentration and purity. | ||
* Followed by digestion and gel electrophoresis, then purification. | |||
* However, concentrations were determined to be too low to complete a ligation. | |||
* More liquid cultures of KAH182 and KAH201 were prepped. | |||
** Colonies placed into LB broth overnight in shaking 37C incubator. | |||
{| class="wikitable" border="0" cellspacing="3" <!-- [DNA] table --> | |||
|- | |||
| <u>Sample</u> || <u>260/280</u> || <u>ng/μL</u> | |||
|- | |||
| 1. KAH201 (Plasmid) || 1.795 || 54.054 | |||
|- | |||
| 2. KAH201 (Plasmid) || 1.708 || 47.244 | |||
|} | |||
{| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table --> | |||
|-valign="top" | |||
| <u>Reagent</u> || <u>Volume</u> || | |||
| rowspan="9" | [[Image:Electrophoresis5.jpg |150px|Results of the fourth electrophoresis attempt]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|- | |||
| DNA (plasmid) || 25 μL | |||
|- | |||
| 10x buffer || 3.0 | |||
|- | |||
| enzyme 1 (Pst1) || 1.0 | |||
|- | |||
| enzyme 2 (Xba1 or Spe1)|| 1.0 | |||
|- | |||
| dH<sub>2</sub>O || 0.0 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) || 0.727 || 2.54 | |||
|- | |||
| 2. KAH182 (X/P) || 0.789 || 1.596 | |||
|} | |||
* Due to the low concentrations, I made more liquid cultures of KAH182 and KAH201. | |||
** Followed the established protocol for making liquid cultures: pick one colony and place into LB broth overnight in shaking 37C incubator. | |||
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Revision as of 12:12, 31 March 2013
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3/25/13
> Measure conc.'s of gel purified fragments
|