User:Karmella Haynes/Notebook/BioBrick cloning/2013/04/12: Difference between revisions
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'''MV9 building: colony PCR''' | '''MV9 building: colony PCR''' | ||
* Pick 8 colonies from plates 1, 2, 3, and 4; make a streak | * Pick 8 colonies from plates 1, 2, 3, and 4; make a mini-streak library, then wash pipette tip in PCR master mix | ||
* Pick 4 colonies from plate 5 (neg. ctrl) | * Pick 4 colonies from plate 5 (neg. ctrl) | ||
* Primers for MV2 vector: DD122B (fwd), DD123B (rev); see [http://partsregistry.org/Part:BBa_J176122 BBa_J176122] | * Primers for MV2 vector: DD122B (fwd), DD123B (rev); see [http://partsregistry.org/Part:BBa_J176122 BBa_J176122] | ||
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| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Volume | | bgcolor=#cfcfcf | Volume | ||
| bgcolor=#cfcfcf | x38 | |||
| rowspan="7" | Expected:<br>MV2 (empty) = 177<br>MV9 (single insert) = 253<br>extra inserts = +76 | | rowspan="7" | Expected:<br>MV2 (empty) = 177<br>MV9 (single insert) = 253<br>extra inserts = +76 | ||
| rowspan="7" | <!-- [[Image:GelImage.jpg|270px|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] --> | | rowspan="7" | <!-- [[Image:GelImage.jpg|270px|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] --> | ||
|- | |- | ||
| DNA (colony) || --- | | DNA (colony) || --- || --- | ||
|- | |- | ||
| 10 μM primer 1 || 1.0 | | 10 μM primer 1 || 1.0 |
Revision as of 19:14, 12 April 2013
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04/12/13
MV9 building: colony PCR
Thermal cycling
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