User:Wesley J. Houston/Notebook/MAP/2013/03/25: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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==3/25/13== | ==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 --> | {| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table --> | ||
|-valign="top" | |-valign="top" | ||
| <u>Reagent</u> || <u>Volume</u> || | | <u>Reagent</u> || <u>Volume</u> || | ||
| rowspan="9" | [[Image:Electrophoresis5.jpg |150px|Results of the fourth electrophoresis attempt]]<br> | | 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 | | DNA (plasmid) || 25 μL | ||
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| || 30 μL --> 37°C/ ~15 min. | | || 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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Latest revision as of 22:34, 26 September 2017
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3/25/13
> Measure conc.'s of gel purified fragments
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