User:Karmella Haynes/Notebook/BioBrick cloning/2013/03/19: Difference between revisions
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== | ==03/19/13== | ||
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* | * Oligo annealing for new mammalian vector | ||
---- | ---- | ||
''' | '''Oligo annealing''' | ||
* Note: made fresh 10x annealing buffer | |||
# New BB 1 | # New BB 1 | ||
* oligo 1 | |||
* oligo 2 | |||
* oligo 3 | |||
* oligo 4 | |||
{| class="wikitable" border="0" cellspacing="3" <!-- Oligo annealing rxn table --> | {| class="wikitable" border="0" cellspacing="3" <!-- Oligo annealing rxn table --> | ||
| DNA (oligos, 100 μM) || | | DNA (oligos, 100 μM) || 4x 3 μL ea. = 12.0 | ||
|- | |- | ||
| 10x annealing buffer || 2.0 | | 10x annealing buffer || 2.0 | ||
|- | |- | ||
| dH<sub>2</sub>O || | | dH<sub>2</sub>O || 6.0 | ||
|- | |- | ||
| || 20 μL | | || 20 μL | ||
|} | |} | ||
Thermal Cycler (try new thermal cycling approach)<br> | |||
Multigene Optimax Thermal Cycler | |||
* 99.9 °C, 10 min. | |||
* [99.9°C, temp. increment -2.0°C, 1 min.; 97.9°C, temp. increment -2.0°C, 1 min.] x35 | |||
* 25°C, ∞ | |||
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Revision as of 17:57, 19 March 2013
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03/19/13
Oligo annealing
Thermal Cycler (try new thermal cycling approach)
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