Haynes Lab:Notebook/Engineering PC-TFs/2015/01/26: Difference between revisions
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==Summary== | ==Summary== | ||
* | '''Ligation Digest and Gel Verification''' | ||
<br> | |||
{| {{table}} border="1" cellspacing="5" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="9" | | |||
<u><b>Expected:</b></u> | |||
<br> | |||
<u>BL05 [XbaI/SpeI]</u><br> | |||
Forward insertion: 3659, 4064 <br> | |||
Reverse insertion: 1538, 6185 <br> | |||
No insertion: 1538, 4064 <br> <br> | |||
<u>CMV/MV9 with [XbaI] </u> <br> | |||
5197<br><br> | |||
| rowspan="10" | [[|400px|Today's gel]]<br> | |||
30μL/lane; 1% agarose;<br> | |||
<u>Observed lengths:</u> <br> | |||
W2 BL05 LCR SV1 (small volume lcr reaction product: 4uL) <br> | |||
W3 BL05 LCR SV2<br> | |||
W4 BL05 LCR LV1 (large volume lcr reaction product: 16uL)<br> | |||
W5 BL05 LCR NEG<br> | |||
[http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|- | |||
| DNA[BL01: W2,W3*](CM9: W4,W5)|| [8.0 μL] (5.0 μL) | |||
|- | |||
| 10X buffer || 3.0 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| SpeI|| 1.0 μL (0uL) | |||
|- | |||
| dH<sub>2</sub>O || [17μL] (21μL) | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br> | |||
Revision as of 11:58, 26 January 2015
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SummaryLigation Digest and Gel Verification
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