Colony PCR

Materials required

Add the following according:

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