IGEM:Peking University/2008/Experiment/PKU 0717 G2 1
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PKU_080717_G2_1
- Screen for pGREF505-lexO clonies
- Plasmid Miniprep-G4M1278-13~24
- Sequencing-G4M1278-14/17/24
- Result for last sequencing: G4M1278-1&2 are right point mutations.
- Point mutation-T1278G/T1743G
- Using right wtGAL4 TG-11
- Point mutation-T1278G/T1743G
- Primer Name:T1743G-F & T1743G-R
- PCR system
| Volume (μL) | |
| 10×buffer iPCR | 5 |
| pGREG505/(TEF-lacI-adhlacS-GFP) | 5 |
| dNTP | 5 |
| KOD Plus | 1 |
| Gal4 template | 1 |
| T1278G-F-Primer | 0.75 |
| T1278G-R-Primer | 0.75 |
| ddH2O | 36.5 |
- conditions
| Temperature(°C) | Time Length(0:00:00) | |
| 94 | 02:00 | |
| 98 | 00:10 | 10 cycles |
| 68 | 00:07 | 30 cycles |
| Hold | 4°C | |
| 98 | 00:10 | 25 cycles |
| 68 | 10:00 | 25 cycles |
| Hold | 4°C |
- Result: have about 7000 band, waiting for sequencing check
- Digestion-point muation product-overnight/4°C
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