User:Karmella Haynes/Notebook/BioBrick cloning/2015/05/05: Difference between revisions
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* Sigma GenElute kit | * Sigma GenElute kit | ||
* Elute w/ 75 μL elution sln. | * Elute w/ 75 μL elution sln. | ||
* Check with | * Check with NotI/XbaI digests | ||
{| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table --> | {| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table --> | ||
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| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Volume | | bgcolor=#cfcfcf | Volume | ||
| rowspan="7" | <u>Expected:</u><br> | | rowspan="7" | <u>Expected:</u><br>KAH87 fwd/MV10 = '''~6150''', 36, 28<br>KAH87 rev/MV10 = ~5100, '''1089''', 28<br>MV10 = ~5100, 36, 28<br> | ||
| rowspan="7" | | | rowspan="7" | [[Image:GelImage.jpg|400px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | ||
|- | |- | ||
| DNA(plasmid) || 2.0 μL | | DNA(plasmid) || 2.0 μL | ||
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| 10X buffer || 1.5 | | 10X buffer || 1.5 | ||
|- | |- | ||
| | | NotI || 1.0 | ||
|- | |- | ||
| | | XbaI || 1.0 | ||
|- | |- | ||
| dH<sub>2</sub>O || 9.5 | | dH<sub>2</sub>O || 9.5 |
Revision as of 16:23, 5 May 2015
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05/05/15
Minipreps
Ryan - PCR & Dig/Lig (promoter insert trial #3) Ryan - Receiver plasmids, assembly - REVISED STRATEGY (PCR promoters)
Thermal cycler: Bio-Rad - Phusion
Cas-tone Project
Minipreps
Assemblies
Oligo annealing
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