User:Karmella Haynes/Notebook/BioBrick cloning/2013/04/01: Difference between revisions
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== | ==04/01/13== | ||
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* | * Check MV9 clones | ||
---- | ---- | ||
'''MV9 clones''' | |||
* Picked colonies from last cloning attempt. Streaked on LB amp agar. | |||
* Grew up 2 mL cultures overnight and did minipreps. | |||
'''Minipreps'''<br> | '''Minipreps'''<br> | ||
* Check with | * Check with S/X digests | ||
** S present in upstream cloning sites, X inside the new insert | |||
{| {{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>1. | | rowspan="7" | <u>Expected:</u><br>1-4. MV9 = 4531, 80 | ||
| rowspan="7" | | | rowspan="7" | [[Image:KAH040113_gel1.jpg|200px|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. | ||
|} | |} | ||
* Result is unclear for #1. Submit for sequencing. | |||
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Revision as of 09:32, 3 April 2013
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04/01/13
MV9 clones
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