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Team:Macquarie Australia/Protocols/PCR
From 2013.igem.org
(Difference between revisions)
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| - | <b>PCR mixture</b> | + | <b><font size = 2>PCR mixture</font></b> |
| - | <br> | + | <br><br> |
| - | 4.0ul 5x phusion buffer | + | 4.0ul 5x phusion buffer<br> |
| - | 0.4ul dNTPs | + | 0.4ul dNTPs<br> |
| - | 0.6ul DMSO | + | 0.6ul DMSO<br> |
| - | 0.2ul Polymerase | + | 0.2ul Polymerase<br> |
| - | 11.8ul water | + | 11.8ul water<br> |
| - | Total 17ul | + | Total 17ul<br> |
| - | 1ul forward primer | + | 1ul forward primer<br> |
| - | 1ul reverse primer | + | 1ul reverse primer<br> |
| - | 1ul template | + | 1ul template<br> |
| + | <br><br> | ||
| + | <b><font size = 2>PCR settings</font></b> | ||
<br><br> | <br><br> | ||
| - | |||
| - | |||
| - | Initial denaturation 98oC for 30 secs | + | Initial denaturation 98oC for 30 secs<br> |
| - | Followed by 30 | + | Followed by 30 repeats of:<br> |
| - | Denaturation 98 oC 10 secs | + | Denaturation 98 oC 10 secs<br> |
| - | Annealing 60 oC 10 secs | + | Annealing 60 oC 10 secs<br> |
| - | Extension 72 oC 2 mins | + | Extension 72 oC 2 mins<br> |
| - | Final extension 72 oC 10 mins | + | Final extension 72 oC 10 mins<br> |
Revision as of 06:48, 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 98oC for 30 secs
Followed by 30 repeats of:
Denaturation 98 oC 10 secs
Annealing 60 oC 10 secs
Extension 72 oC 2 mins
Final extension 72 oC 10 mins
