Team:Macquarie Australia/Protocols/PCR

From 2013.igem.org

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<center><h1>Polymerase Chain Reaction</h1></center>
<center><h1>Polymerase Chain Reaction</h1></center>
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[[File:Thermocycler.JPG|350px|thumb|left|Thermocycler]]
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<b>PCR mixture</b>
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<b><font size = 2>PCR mixture</font></b>
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<br>
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<br><br>
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     4.0ul 5x phusion buffer
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     4.0ul 5x phusion buffer<br>
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     0.4ul dNTPs
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     0.4ul dNTPs<br>
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     0.6ul DMSO
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     0.6ul DMSO<br>
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     0.2ul Polymerase
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     0.2ul Polymerase<br>
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     11.8ul water
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     11.8ul water<br>
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     Total 17ul
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     Total 17ul<br>
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     1ul forward primer
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     1ul forward primer<br>
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     1ul reverse primer
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     1ul reverse primer<br>
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     1ul template
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     1ul template<br>
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<br><br>
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<b>PCR settings</b>
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<b><font size = 2>PCR settings</font></b>
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<br>
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<br><br>
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     Initial denaturation 98oC for 30 secs
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     Initial denaturation 98°C for 30 secs<br>
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     Followed by 30 repears of:
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     <b>Followed by 30 repeats of:</b><br>
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         Denaturation 98 oC 10 secs
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         <b>Denaturation -</b> 98 °C for 10 secs<br>
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         Annealing 60 oC       10 secs
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         <b>Annealing -</b> 60°C       for 10 secs<br>
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         Extension 72 oC       2 mins
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         <b>Extension -</b> 72°C       for 2 mins<br>
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         Final extension 72 oC 10 mins
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         <b>Final extension -</b> 72°C for 10 mins<br>

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

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