Team:Macquarie Australia/Protocols/PCR

From 2013.igem.org

(Difference between revisions)
Line 22: Line 22:
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<br><br>
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     Initial denaturation 98oC for 30 secs<br>
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     Initial denaturation 98°C for 30 secs<br>
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     Followed by 30 repeats of:<br>
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     <b>Followed by 30 repeats of:</b><br>
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         Denaturation 98 oC 10 secs<br>
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         Denaturation 98 °C 10 secs<br>
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         Annealing 60 oC       10 secs<br>
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         Annealing 60°C       10 secs<br>
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         Extension 72 oC       2 mins<br>
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         Extension 72°C       2 mins<br>
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         Final extension 72 oC 10 mins<br>
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         Final extension 72°C 10 mins<br>

Revision as of 06:50, 13 September 2013


Polymerase Chain Reaction


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 10 secs
Annealing 60°C 10 secs
Extension 72°C 2 mins
Final extension 72°C 10 mins

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