Team:EPF-Lausanne/Protocols/1.5 steps PCR

From 2009.igem.org

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! scope=row | 5            
! scope=row | 5            
|Cycle 2-4 10 times
|Cycle 2-4 10 times
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|-
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! scope=row | 6            
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|72°C
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|7:00
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|-
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! scope=row | 7            
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|4°C
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|∞
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</center>
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'''3.''' We now add the standard Igem primer which finish the extension, and do the following cycles:
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<center>
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{| class="wikitable" style="text-align:left; width:20%;"
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|-
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! scope=row | 1&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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|94°C
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|4:00
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! scope=row | 2&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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|94°C
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|0:30
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|-
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! scope=row | 3&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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|55°C
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|1:00
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|-
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! scope=row | 4&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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|72°C
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|2:00
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! scope=row | 5&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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|Cycle 2-4 25-30 times
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Revision as of 09:48, 4 September 2009



Miniprep Protocol




1. Do the following mixing:

dNTP 1 μl
5x Buffer + MgCl2 10 μl
Primer Mix 1 μl
template 0.5 μl
HiFi Plus DNAp 0.5 μl
dH2O 36.5 μl
Final Volume 50 μl


2. Make it run 10 cycles:

1             94°C 4:00
2             94°C 0:30
3             55°C 1:00
4             72°C 2:00
5             Cycle 2-4 10 times
6             72°C 7:00
7             4°C ∞


3. We now add the standard Igem primer which finish the extension, and do the following cycles:

1             94°C 4:00
2             94°C 0:30
3             55°C 1:00
4             72°C 2:00
5             Cycle 2-4 25-30 times
6             72°C 7:00
7             4°C ∞


Retrieved from "http://2009.igem.org/Team:EPF-Lausanne/Protocols/1.5_steps_PCR"