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Team:Newcastle/Colony PCR
From 2010.igem.org
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Colony PCR
Materials required
Add the following according:
- 37.5 µl of distilled H2O
- 10 µl of 5x GoTaq Buffer
- Nucleotide DNTP
- 2.5 µl forward primer
- 2.5 µl backward primer
- 1 µl template DNA
Conditions for ThermoCycler:
- Initialise - 95°C for 2 minutes.
- Denature - 95°C for 30 seconds.
- Anneal - 52°C for 30 seconds (melting temperature, Tm, of template)
- Extension - 75°C for 30 seconds
- Extension finish - 75°C for 5 minutes
- Hold - 4°C
Steps 2 to 4 are repeated for 30 cycles before continuing to step 5.
   
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