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 PcTF Subcloning in E. coli
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3/5/2013
PcTF + His Tag Ligation in V0120
PCR
- 95°C, 3 min.
- [95°C, 30 sec.; 57°C, 30 sec.; 72°C, 1 min.] x35
- 72°C, 3 min.
- 4°C, ∞
| Reagent
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Volume
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Expected:
1. KAH109 = 1132
2. KAH111 = 1132
3. KAH112 = 1132
4. KAH115 = 931
5. KAH148 = 967
6. KAH150 = 967
7. KAH151 = 967
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10 μL/lane; 1% agarose; Ladder
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| DNA(plasmid) |
0.2 μL
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| 10 μM primer 1 |
1.0
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| 10 μM primer 2 |
1.0
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| 2x GoTaq |
25.0
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| dH2O |
22.8
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50.0 μL
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Purification
- Clean with Zymo DNA c&c; elute w/ 20 μL dH2O
- Measure [DNA]
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PcTF Subcloning in E. coli
