User:Daniel A. Vargas/Notebook/General lab notebook/2015/08/04: Difference between revisions
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* Incubate at 37°C/ 30 min. | * Incubate at 37°C/ 30 min. | ||
* Heat-inactivate PNK at 65°C/ 20 min. | * Heat-inactivate PNK at 65°C/ 20 min. | ||
LCR Reaction | LCR Reaction | ||
* Add components to the PNK DNA reaction as described in the table below | |||
* Set up the reactions in PCR-sized tubes | |||
{| {{table}} | |||
| Reagent || Volume | |||
|- | |||
| PNK DNA || 10 μL | |||
|- | |||
| Oligo Bridge || 3.0 (each) | |||
|- | |||
| 10X Ampligase Buffer || 3.0 | |||
|- | |||
| Ampligase || 1.0 | |||
|- | |||
| dH<sub>2</sub>O || x μL | |||
|- | |||
| || 30.0 μL | |||
|} | |||
* Run LCR... | |||
** Look for "LCR" program on one of the PCR machines | |||
Transformation | |||
* Carry out the usual transformation. (quick and dirty.) | |||
Revision as of 22:26, 3 August 2015
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ASSEMBLY of MV10, ATF2, and Gal4DB/mCherry
LCR Assembly Prep the Bridge Oligos
|