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From 2010.igem.org

Colony PCR

Materials to add accordingly:

1. 37.5µl of distilled H2O
2. 10µl of 5x GoTaq Buffer
3. Nucleotide DNTP
4. 2.5µl forward primer
5. 2.5µl backward primer
6. 1µl template DNA

Conditions for ThermoCycler:

1. Initialise - 95°C for 2 minutes.
2. Denature - 95°C for 30 seconds.
3. Anneal - 52°C for 30 seconds (Melting temperature, Tm of template).
4. Extension - 75°C for 30 seconds
5. Extension finish - 75°C for 5 minutes
6. Hold - 4°C

Steps 2 to 4 are repeated for 30 cycles before continuing to step 5.