Team:Groningen/protocols/Colony PCR

From 2013.igem.org

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Revision as of 08:48, 19 September 2013

Colony PCR

Materials:
  • PCR tubes
  • MQ water
  • DMSO 5%
  • Phusion buffer
  • DNTPs
  • F- and R-primer
  • Phusion Polymerase
  • DNA template


PCR reaction mixture:
Component 20µl Final concentration
MQ water up to 20µl
5x Phusion Buffer 2µl 1x
DMSO 5% 1.5µl
10mM dNTPs 1µl 200µM
primer F 1µl 0.5µM
primer R 1µl 0.5µM
template DNA 1µl ∼1ng
Phusion Polymerase
2U/µl		 0.3µl 0.01U/µl


Cycling instructions:
Temperature Time Number of Cycles
98°C 10min 1
98°C 10sec 34
Tm-5°C 25sec
72°C 30sec/kb
72°C 10min 1
4°C ∞ 1