User:Karmella Haynes/Notebook/BioBrick cloning/2012/12/11: Difference between revisions
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(Autocreate 2012/12/11 Entry for User:Karmella_Haynes/Notebook/BioBrick_cloning) |
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== | ==12/11/12== | ||
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* | * Assembly: KAH111 + C-His (trouble shooting for Behzad, start from plasmids) | ||
* | * Minipreps: Rene's pSB1A3-RFP (2); cut with X/S for Brady and Vi's Gibson cloning | ||
---- | ---- | ||
'''Minipreps'''<br> | '''Minipreps'''<br> | ||
* | * pSB1A3-RFP (retransformation from 12/10/12) | ||
* Concentrations: (1) 260 = , 260/280 = , '''ng/μL = '''; (2) 260 = , 260/280 = , '''ng/μL = '''; | |||
* Digest 10 μL of each with X/S for Gibson cloning (Brady and Vi) | |||
{| {{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>3. pSB1A3 = size<br>4. BB2 = size<br> | ||
| rowspan="7" | | | rowspan="7" | [[Image:KAH121112_gel1.png|400px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | ||
|- | |- | ||
| DNA(plasmid) || | | DNA(plasmid) || 10.0 μL | ||
|- | |- | ||
| 10X buffer || 1.5 | | 10X buffer || 1.5 |
Revision as of 16:20, 11 December 2012
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12/11/12
Minipreps
Assemblies
Oligo annealing
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