User:Karmella Haynes/Notebook/BioBrick cloning/2015/04/16: Difference between revisions
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| bgcolor=#cfcfcf | Rxn1,3 | | bgcolor=#cfcfcf | Rxn1,3 | ||
| bgcolor=#cfcfcf | Rxn2,4 | | bgcolor=#cfcfcf | Rxn2,4 | ||
| rowspan="7" | Expected:<br>MV10 band = | | rowspan="7" | Expected:<br>MV10 band = 5191 | ||
| rowspan="7" | [[Image:KAH041615_gel1.jpg|250px|Hover name]]<br>10 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | | rowspan="7" | [[Image:KAH041615_gel1.jpg|250px|Hover name]]<br>10 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | ||
|- | |- | ||
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* 72°C, 10 min | * 72°C, 10 min | ||
* 4°C ∞ | * 4°C ∞ | ||
Conclusion | |||
* Not ideal, proceed with caution | |||
Measure conc. | |||
* DNA purified with Sigma PCR clean-up kit | |||
* Eluted& back-eluted with 30 μL elution sln. | |||
{| {{table}} cellspacing="3" <!-- [DNA] table --> | |||
|- bgcolor=#cfcfcf | |||
| Sample || OD260 || 260/280 || ng/μL | |||
|- | |||
| 1. 5'p-MV10 PCR || ### || ### || ### | |||
|} | |||
Revision as of 16:49, 16 April 2015
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04/16/15
LCR Development
Measure conc.
Program: Phusion (block B)
Conclusion
gRNA cloning - Rene
Phosphorylate and anneal oligo pairs
Bio-Rad Thermocycler
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