Haynes Lab:Notebook/Engineering PC-TFs/2014/08/01: Difference between revisions
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==Summary== | ==Summary== | ||
'''Ligation Digest and Gel Verification (2)''' | |||
<br> | |||
{| {{table}} border="1" cellspacing="5" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="9" | | |||
<u>Expected:</u> | |||
<br> | |||
BL09 Ligation Colonies 1-3[XbaI/NruI] (Well: 2-4)<br> | |||
Forward insertion: 710, 5921 <br> | |||
Reverse insertion: 2141, 4490 <br> | |||
No insertion: 710, 4490<br> <br> | |||
CMV/MV9 with [XbaI/NruI] (Well: 5) <br> | |||
710, 4490<br><br> | |||
CMV/MV9 [XbaI](Well: 6) <br> | |||
5200<br><br> | |||
Dephos CMV/MV9 (Well: 7) <br> | |||
5200 | |||
| rowspan="10" | [[Image:20140801 132828.jpg|400px|Today's gel]]<br> | |||
15 μL/lane; 1% agarose;<br> | |||
<u>Observed lengths:</u> <br> | |||
W2(BL09Lig1): 5000, 7100, 2500; <br> | |||
W3(BL09Lig2): 5000, 7100, 2500; <br> | |||
W4(BL09Lig3): 5000, 7100, 2500; <br> | |||
W5(Empty CM9): 5000, 700; <br> | |||
W6(CM9XbaI): 10000, 20000, 5000.<br> | |||
W7(DephosCM9): 5000 <br> | |||
[http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|- | |||
| DNA(BL01x2, BL05x1, BL09x2, CMV/MV9x1) || 3.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| NruI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 8.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br> | |||
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__NOTOC__ | __NOTOC__ |
Revision as of 12:53, 3 August 2014
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SummaryLigation Digest and Gel Verification (2)
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