User:Karmella Haynes/Notebook/BioBrick cloning/2013/01/11: Difference between revisions
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(Autocreate 2013/01/11 Entry for User:Karmella_Haynes/Notebook/BioBrick_cloning) |
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== | ==01/11/13== | ||
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* | * Golden Gate assembly trial 1: mini preps/ digests | ||
* | * Golden gate assembly trial 2: transformation | ||
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| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Volume | | bgcolor=#cfcfcf | Volume | ||
| rowspan="7" | <u>Expected:</u><br>1. | | rowspan="7" | <u>Expected:</u><br>1-4. hPCD-BL01/pSB1A3 = 2686, 2000 | ||
| rowspan="7" | | | rowspan="7" | [[Image:GelImage.jpg|300px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | ||
|- | |- | ||
| DNA(plasmid) || | | DNA(plasmid) || 3.0 μL | ||
|- | |- | ||
| 10X buffer || 1.5 | | 10X buffer || 1.5 | ||
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| PstI || 1.0 | | PstI || 1.0 | ||
|- | |- | ||
| dH<sub>2</sub>O || | | dH<sub>2</sub>O || 8.5 | ||
|- | |- | ||
| || 15 μL --> 37°C/ ~15 min. | | || 15 μL --> 37°C/ ~15 min. | ||
|} | |} | ||
* None of the digests show both expected bands; probably just vector | |||
* | |||
---- | ---- | ||
''' | '''Golden gate assembly trial 2''' | ||
* Add 10.0 μL assembly reaction to 50.0 μL DH5α-Turbo; ice/ 5 min.; plate on amp agar | |||
Revision as of 15:04, 11 January 2013
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01/11/13
Minipreps
Golden gate assembly trial 2
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