Team:Macquarie Australia/Protocols/PCR

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[[File:Thermocycler.JPG|400px|thumb|left|Thermocycler]]
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[[File:Thermocycler.JPG|350px|thumb|left|Thermocycler]]

Latest revision as of 22:33, 17 September 2013


Polymerase Chain Reaction


Thermocycler


PCR mixture

4.0ul 5x phusion buffer
0.4ul dNTPs
0.6ul DMSO
0.2ul Polymerase
11.8ul water
Total 17ul
1ul forward primer
1ul reverse primer
1ul template


PCR settings

Initial denaturation 98°C for 30 secs
Followed by 30 repeats of:
Denaturation - 98 °C for 10 secs
Annealing - 60°C for 10 secs
Extension - 72°C for 2 mins
Final extension - 72°C for 10 mins