User:Karmella Haynes/Notebook/BioBrick cloning/2015/05/22: Difference between revisions
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* Digests | * Digests | ||
** Check with EcoRI digests - promoter insertion generates a new EcoRI site | ** Check with EcoRI digests - promoter insertion generates a new EcoRI site | ||
** Note: BraR = 776, pBra = 214 | |||
{| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table --> | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1-8. pBra-BraR_MRV = 2989, 1202<br>empty vector = 3977<br> | |||
| 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 | |||
|- | |||
| 10X buffer || 1.5 | |||
|- | |||
| EcoRI || 1.0 | |||
|- | |||
| dH<sub>2</sub>O || 10.5 | |||
|- | |||
| || 15 μL --> 37°C/ ~15 min. | |||
|} | |||
---- | |||
'''Cas-tone - H3_MV11 minipreps & digests''' | |||
* Cultures | |||
** Pick 2 colonies from each ligation plate | |||
** Make streak library & 5 mL cultures | |||
* Minipreps<br> | |||
** Sigma GenElute kit, elute w/ 75 μL elution sln. | |||
* Digests | |||
** Check with ### digests - promoter insertion generates a new EcoRI site | |||
** Note: BraR = 776, pBra = 214 | ** Note: BraR = 776, pBra = 214 | ||
Revision as of 11:11, 22 May 2015
Karmella's BioBrick Cloning | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
05/22/15
Ryan - Bra minipreps & digests
Cas-tone - H3_MV11 minipreps & digests
Assemblies
Oligo annealing
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