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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> | + | <b><font size = 2>PCR mixture</font></b> |
| - | <br> | + | <br><br> |
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| + | 4.0ul 5x phusion buffer<br> | ||
| + | 0.4ul dNTPs<br> | ||
| + | 0.6ul DMSO<br> | ||
| + | 0.2ul Polymerase<br> | ||
| + | 11.8ul water<br> | ||
| + | Total 17ul<br> | ||
| + | 1ul forward primer<br> | ||
| + | 1ul reverse primer<br> | ||
| + | 1ul template<br> | ||
| + | <br><br> | ||
| - | + | <b><font size = 2>PCR settings</font></b> | |
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<br><br> | <br><br> | ||
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| - | Initial denaturation | + | Initial denaturation 98°C for 30 secs<br> |
| - | Followed by 30 | + | <b>Followed by 30 repeats of:</b><br> |
| - | Denaturation 98 | + | <b>Denaturation -</b> 98 °C for 10 secs<br> |
| - | Annealing | + | <b>Annealing -</b> 60°C for 10 secs<br> |
| - | Extension | + | <b>Extension -</b> 72°C for 2 mins<br> |
| - | Final extension | + | <b>Final extension -</b> 72°C for 10 mins<br> |
Latest revision as of 22:33, 17 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 for 10 secs
Annealing - 60°C for 10 secs
Extension - 72°C for 2 mins
Final extension - 72°C for 10 mins
