Haynes Lab:Notebook/Engineering PC-TFs/2014/09/01: Difference between revisions
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==Summary== | ==Summary== | ||
* | * Split cells. | ||
* Restriction digest/gel verification of PcTF CMV/MV9 ligation. | |||
---- | |||
''''Restriction Digest/Gel Verification'''' | |||
{| {{table}} border="1" cellspacing="5" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="9" | | |||
<u><b>Expected:</b></u> | |||
<br> | |||
<br>BL01(Well: 4-8, 10-13), BL05(Well:14) <br> | |||
Forward insertion: 710, 7010. <br> | |||
Reverse insertion: 3230, 4490. <br> | |||
No insertion: 710, 4490. <br><br> | |||
<u>CMV/MV9 [XbaI/NruI] (Well: 9)</u> <br> | |||
710, 4490<br><br> | |||
| rowspan="10" | [[IMAGE URL HERE|400px|Today's gel]]<br> | |||
15 μL/lane; 1% agarose;<br> | |||
<u>Observed lengths:</u> <br> | |||
W2(BL01Lig1): 5000, 700; <br> | |||
W3(BL05Lig1): 5000, 700; <br> | |||
W4(BL05Lig2): 5000, 700; <br> | |||
W5(BL05Lig3): 5000, 700; <br> | |||
W6(BL05Lig4): 5000, 700; <br> | |||
W7(Empty CMV/MV9): 5000, 700; <br> | |||
W8(BL09Lig1): 5000, 700; <br> | |||
W9(BL09Lig2): 7000, 700; <br> | |||
W10(BL09Lig3): 5000, 700; <br> | |||
W11(BL09Lig4): 5000, 700; <br> | |||
W12(BL09Lig5): 7000, 700;<br> | |||
W13(BL09Lig6): 5000, 700; <br> | |||
[http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|- | |||
| DNA(BL01x2, BL05x1, BL09x2, CMV/MV9x1) || 2.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| NruI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 9.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
Revision as of 16:46, 1 September 2014
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Summary
'Restriction Digest/Gel Verification'
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