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 Karmella's BioBrick Cloning
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03/19/13
- Oligo annealing for new mammalian vector based on MV9
Oligo annealing
- Note: made fresh 10x annealing buffer
- MV6(X/S) + K-XbaI-NLS-his-stop
- K-XbaI-NLS-his-st 1
- K-XbaI-NLS-his-st 2
- K-XbaI-NLS-his-st 3
- K-XbaI-NLS-his-st 4
| DNA (oligos, 100 μM) |
4x 3 μL ea. = 12.0
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| 10x annealing buffer |
2.0
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| dH2O |
6.0
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20 μL
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Thermal Cycling (try new thermal cycling approach)
Multigene Optimax Thermal Cycler
- 99.9 °C, 10 min.
- [99.9°C, temp. increment -2.0°C, 1 min.; 90.0°C, temp. increment -2.0°C, 1 min.] x35
- 25°C, ∞
Vector Digest
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Karmella's BioBrick Cloning
