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Team:Newcastle/Colony PCR
From 2010.igem.org
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# Initialise - 95°C for 2 minutes. | # Initialise - 95°C for 2 minutes. | ||
# Denature - 95°C for 30 seconds. | # Denature - 95°C for 30 seconds. | ||
| - | # Anneal - 52°C for 30 seconds (melting temperature, Tm, of template) | + | # Anneal - 52°C for 30 seconds (melting temperature, Tm, of template) |
# Extension - 75°C for 30 seconds | # Extension - 75°C for 30 seconds | ||
# Extension finish - 75°C for 5 minutes | # Extension finish - 75°C for 5 minutes | ||
Revision as of 09:16, 27 July 2010
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Colony PCR
Materials to add accordingly:
- 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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