User:Wesley J. Houston/Notebook/MAP/2013/03/07
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* Prepped plasmids using Zyppy miniprep kit. | * Prepped plasmids using Zyppy miniprep kit. | ||
| - | * Digested plasmids with EcoR1 and Pst1 | + | * Digested plasmids with EcoR1 and Pst1. |
| - | * Ran gel electrophoresis to confirm assembly ( | + | * Ran gel electrophoresis to confirm assembly. |
| - | * No DNA present, repeat ligation. | + | |
| + | {| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table --> | ||
| + | |-valign="top" | ||
| + | | <u>Reagent</u> || <u>Volume</u> || | ||
| + | | rowspan="9" | [[Image:Electrophoresis3.jpg |150px|Results of the ligation electrophoresis]]<br>15 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 || 1.0 | ||
| + | |- | ||
| + | | enzyme 2 || 1.0 | ||
| + | |- | ||
| + | | dH<sub>2</sub>O || 5.0 μL | ||
| + | |- | ||
| + | | || 30 μL --> 37°C/ ~15 min. | ||
| + | |} | ||
| + | |||
| + | |||
| + | * No DNA present, will repeat ligation. | ||
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | ||
Revision as of 16:17, 18 March 2013
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