Haynes Lab:Notebook/Engineering PC-TFs/2014/09/01: Difference between revisions
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|style="background-color: #FFCC00"|<br>[[Image:Hayneslab3.gif|200px|center]][[Image:Asu logo 3.gif|200px|center]]<span style="font-size:22px;"><br></span> | |style="background-color: #FFCC00"|<br>[[Image:Hayneslab3.gif|200px|center]][[Image:Asu logo 3.gif|200px|center]]<span style="font-size:22px;"><br></span> | ||
|style="background-color: #800000" align="center"| | |style="background-color: #800000" 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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==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, 11-14), BL05(Well:2) <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:20140901 180224.jpg|400px|Today's gel]]<br> | |||
15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]<br> | |||
<u>Observed lengths:</u> <br> | |||
W2(BL05Lig1): 5000, 700; <br> | |||
W4(BL01Lig1): 3200, 4500; <br> | |||
W5(BL01Lig2): 5000, 700; <br> | |||
W6(BL01Lig3): 5000, 700; <br> | |||
W7(BL01Lig4): 5000, 700; <br> | |||
W8(BL01Lig5): 5000, 700; <br> | |||
W9(Empty CMV/MV9): 5000, 700; <br> | |||
W11(BL01Lig6): 5000, 700; <br> | |||
W12(BL01Lig7): 5000, 700; <br> | |||
W13(BL01Lig8): 5000, 700;<br> | |||
W14(BL01Lig9): 5000, 700; <br> | |||
|- | |||
| 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. | |||
|} | |||
<br> | |||
One reverse insertion. No forward insertions. Will reattempt ligation. | |||
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Latest revision as of 00:13, 27 September 2017
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Summary
Restriction Digest/Gel Verification
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