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Engineering PC-TFs
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01/25/13
- Type IIS assembly - day 1
PCR
- Resuspend new primers to a final concentration of 100 μM
- Make 10 μM dilutions in separate tubes
- All of the reactions are as follows:
- template, primer 1, primer 2
- template, primer 1, primer 2
- template, primer 1, primer 2
- template, primer 1, primer 2
Reagent
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Volume
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Expected: 1. pSB1A3 = ~2000 2. hPCD = 186 3. BL01 = 2520
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10 μL/lane; 1% agarose; Ladder
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DNA(plasmid) |
0.2 μL
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primer 1 |
1.0
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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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PCR
- 95°C, 3 min.
- [95°C, 30 sec.; 57°C, 30 sec.; 72°C, 1 min.] x35
- 72°C, 3 min.
- 4°C, ∞
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