Team:SDU-Denmark/labwork/Protocols/PCR

From 2012.igem.org

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H2O up to 50μl </br>
H2O up to 50μl </br>
</Br>
</Br>
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<b>PCR-PROGRAM:</b> </br>
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<b>PCR program: </b></br>
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<pre>
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<pre>95°C for 2 minutes
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1. Start 95°C 2min </br>
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<pre>29 cycles:
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2. Denaturing 95°C 1min </br>
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95°C for 1 minut 
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3. Annealing 55°C 30sek</br>
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55°C for 30 sek
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4. Elonging 72°C 1min (1kb/min) </br>
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72°C for 1min (1kb/min)</pre>
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5. GO TO step 2, x 29 cycles</br>
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72°C for 5 minutes </br>
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6. End 72°C 5min</br>
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4°C on hold</pre>
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7. Hold 4°C infinite </br>
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</pre>
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Revision as of 14:04, 22 September 2012

iGEM TEAM ::: SDU-DENMARK courtesy of NIAID


mRNA Isolation PCR Miniprep Check Digest
3A-Assembly Colony-PCR Transformation Gel-electrophoresis
Reverse Transcriptase Mutagenisis PCR-,gel clean-up Content

Polymerase Chain Reaction

Proof-reading PCR using Phusion Hot Start II


some_text

PCR program: 
95°C for 30 seconds 

25-35 cycles:
98°C for 30 seconds 
Annealing temperature (5°C below primer melting temperature)  
72°C for 15-30s/kb

72°C for 5-10 minutes 

4°C on hold


Non-proof-reading PCR using Taq Polymerase


5μl Dream Taq Buffer
5μl dNTP
1μl VF2
1μl VR
Udtag 10μl fra primer stock og fortynd i 90μl oprenset vand
1,5μl Dream Taq Polymerase
1μl Template
H2O up to 50μl

PCR program: 
95°C for 2 minutes 

29 cycles:
95°C for 1 minut   
55°C for 30 sek
72°C for 1min (1kb/min)

72°C for 5 minutes 

4°C on hold

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