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Team:Chiba/protocol/PCR
From 2008.igem.org
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PCR | PCR | ||
| + | |||
| + | *Resuspend primer in Nuculease free water to 100 µM | ||
| + | |||
| + | *PCR mix | ||
| + | #DNA template 1µL | ||
| + | #Fwd primer 10µL (final con. 10 pM) | ||
| + | #Rev primer 10µL | ||
| + | #10x thermo pol buffer 10µL | ||
| + | #dNTP mix 10µL | ||
| + | #DNA pol. 1µL | ||
| + | #dH20 58µL | ||
| + | <br>---------------------------- | ||
| + | <br> 100µL | ||
| + | |||
| + | |||
| + | *PCR cycle | ||
| + | |||
| + | Start: 94 °C for 5 min. (melt) | ||
| + | cycle: melt: 1 min. | ||
| + | anneal : 30 sec. | ||
| + | cycle end: extension:72 °C for 3.5 min. | ||
| + | 25 cycles | ||
| + | 72 °C for 10 min | ||
| + | store: keep at 6 °C forever | ||
Latest revision as of 02:13, 11 September 2008
PCR
- Resuspend primer in Nuculease free water to 100 µM
- PCR mix
- DNA template 1µL
- Fwd primer 10µL (final con. 10 pM)
- Rev primer 10µL
- 10x thermo pol buffer 10µL
- dNTP mix 10µL
- DNA pol. 1µL
- dH20 58µL
----------------------------
100µL
- PCR cycle
Start: 94 °C for 5 min. (melt)
cycle: melt: 1 min.
anneal : 30 sec.
cycle end: extension:72 °C for 3.5 min.
25 cycles
72 °C for 10 min
store: keep at 6 °C forever
