Haynes Lab:Notebook/Engineering PC-TFs/2013/09/12: Difference between revisions
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== | ==September 12, 2013== | ||
* Follow steps for gel electrophoresis. | * Follow steps for gel electrophoresis. | ||
* Orient top notch of gel electrophoresis apparatus. Place gel tray into larger tray. | * Orient top notch of gel electrophoresis apparatus. Place gel tray into larger tray. | ||
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| bgcolor=#cfcfcf | '''Reagent''' | | bgcolor=#cfcfcf | '''Reagent''' | ||
| bgcolor=#cfcfcf | '''Volume''' | | bgcolor=#cfcfcf | '''Volume''' | ||
| rowspan="7" | [[Image: | | rowspan="7" | [[Image:cmvgel2.jpg|250px| CMV and MV9 Stock digest]] [[Image:cmvgel1_cutout.jpg|250px|CMV and MV9 Stock Digest 3/18/2013]] <br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | ||
|- | |- | ||
| DNA (plasmid) || 20.0 μL | | DNA (plasmid) || 20.0 μL | ||
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|- | |- | ||
| || 30 μL --> 37°C/ ~10 min. | | || 30 μL --> 37°C/ ~10 min. | ||
|} | |||
''Zymoclean<sup>TM</sup> Gel DNA Recovery Kit'' | |||
*Add 3 volumes of ADB Buffer to each volume of gel (600 μL). | |||
*Incubate at 55°C for 10 minutes. | |||
*Centrifuge at ≥ 10,000 x g for 30 seconds. Discard flow-through. | |||
*Add 200 μL of DNA Wash Buffer to the column and centrifuge for 30 seconds. Repeat wash step. | |||
*Place Zymoclean Column into a new 1.5 mL tube. Add 20μL of dH<sub>2</sub>O to elute DNA | |||
{|border="1" cellpadding="5" cellspacing="0" align="left" | |||
|- | |||
! scope="col" style="background:#efefef;" | # | |||
! scope="col" style="background:#efefef;" | Sequence | |||
! scope="col" style="background:#efefef;" | ng/µL | |||
|- | |||
|1 | |||
|CMV | |||
| -1.17 | |||
|- | |||
|2 | |||
|CMV | |||
|2.767 | |||
|- | |||
|3 | |||
|CMV | |||
| -2.116 | |||
|- | |||
|5 | |||
|MV9 | |||
|20.838 | |||
|- | |||
|} | |} | ||
Revision as of 21:19, 11 October 2013
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September 12, 2013
ZymocleanTM Gel DNA Recovery Kit
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