Team:Newcastle/Colony PCR

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(Colony PCR)
 
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=='''Colony PCR'''==
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=Colony PCR=
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====Materials to add accordingly:====
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==Materials required==
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Add the following according:
# 37.5 µl of distilled H2O
# 37.5 µl of distilled H2O
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# 1 µl template DNA
# 1 µl template DNA
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==Conditions for ThermoCycler:==
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====Conditions for ThermoCycler:====
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# Initialise - 95°C for 2 minutes.
# Initialise - 95°C for 2 minutes.

Latest revision as of 14:52, 28 July 2010

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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.

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