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Team:Paris Bettencourt/Notebook/Phage Sensor/Monday 23rd September.html
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| - | <b>Gel<b><br> | + | <b>Gel</b><br> |
100V, 1%, 1,5h<br> | 100V, 1%, 1,5h<br> | ||
<br> | <br> | ||
Picture: 13/09/23_SPCR16_17_corresponding primers<br> | Picture: 13/09/23_SPCR16_17_corresponding primers<br> | ||
<br> | <br> | ||
| - | <b>PCR<b><br> | + | <b>PCR</b><br> |
SPCR 16 (normal phusion at 48°C)<br> | SPCR 16 (normal phusion at 48°C)<br> | ||
<br> | <br> | ||
Revision as of 12:49, 2 October 2013
Detect
ASDFMonday 23rd September
Gels and PCR
Gel100V, 1%, 1,5h
Picture: 13/09/23_SPCR16_17_corresponding primers
PCR
SPCR 16 (normal phusion at 48°C)
| Reagent | Volume |
| x1 | |
| Nuclease-free water | 37.25 µl |
| 5x Phusion HF Buffer | 10 µl |
| 10 mM dNTPs | 1 µl |
| Forward Primer (10 uM) | 0.5 µl |
| Reverse Primer (10 uM) | 0.5 µl |
| Template Plasmid | 0.25 µl |
| Phusion DNA Polymerase | 0.5 µl |
| Total Volume | 50 µl |
| Thermocycler Protocol: NEB Phusion | ||||
| Temp | Time | |||
| Start | 98°C | 30 sec | Melt | |
| Cycle 1 | 98°C | 5 sec | Melt | 35 cycles |
| Cycle 2 | 72°C | 30 sec | Anneal | |
| Cycle 3 | 72°C | 30 sec per kb | Extend | |
| Finish | 72 °C | 5 min | Extand | |
| Store | 10°C | Forever | Store |
