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 Project name
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4/1/13
- Completed 2 ligations of KAH201 (S/P)/3403 + KAH182 (X/P)/1675
- First ligation used KAH201 (S/P)/3403 and KAH182 (X/P) 1675 made on 3/26/13
- Second ligation used the same KAH182 (X/P) 1675 and KAH201 (S/P)/3403 made on 2/8/13
> Ligation 1
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1 |
2 |
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| Insert DNA (KAH182) |
3.5 |
--- |
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| Vector DNA (KAH201) |
1.0 |
1.0 |
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| 2x lgn buf (Roche) |
5.5 |
5.5 |
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| T4 ligase (NEB) |
1.0 |
1.0 |
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| dH2O |
0.0 |
3.5 |
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11 μL |
11 μL |
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- Follow quick transformation protocol
- Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar
| Ligation |
Plate results (lig : neg crtl)04/01/13: Ligation 1
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| 1. KAH201 (S/P)/3403, 25 ng + KAH182 (X/P)/1675, 26 ng |
0
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| 2. KAH201 (S/P)/3403, 25 ng |
0
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> Ligation 2
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1 |
2 |
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| Insert DNA (KAH182) |
3.5 |
--- |
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| Vector DNA (KAH201) |
0.7 |
0.7 |
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| 2x lgn buf (Roche) |
5.2 |
5.2 |
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| T4 ligase (NEB) |
1.0 |
1.0 |
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| dH2O |
0.6 |
4.1 |
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11 μL |
11 μL |
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- Follow quick transformation protocol
- Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar
| Ligation |
Plate results (lig : neg crtl)04/01/13: Ligation 2
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| 1. KAH201 (S/P)/3403, 25 ng + KAH182 (X/P)/1675, 26 ng |
~40
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| 2. KAH201 (S/P)/3403, 25 ng |
0
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- Ligation 2 appears successful, with ~40 colonies in the ligation plate and 0 colonies in the negative control plate.
- The next step will be to confirm the assembly, to be completed on 4/4/13
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