Team:TU Darmstadt/protocols/PCR

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<B> Chemicals & consumables <br></B>
<B> Chemicals & consumables <br></B>
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DNA tamplate<br>
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DNA template<br>
Nuclease-free water<br>
Nuclease-free water<br>
Primer<br>
Primer<br>
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<br>
<B> Procedure<br></B>
<B> Procedure<br></B>
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This is a commonly used protocol thta we used for Taq-polymerase<br>
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PCR program (30 cycles)
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<B>PCR program (30 cycles)<br></B>
initial denaturation 95°C, 60s
initial denaturation 95°C, 60s
denaturation 95°C, 45s
denaturation 95°C, 45s
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annealing 54,5°C, 90s
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annealing 54,5°C, 90s  
elongation 72°C, 120s
elongation 72°C, 120s
final elongation 72°C, 120s
final elongation 72°C, 120s
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run on an analytic 1% agarose gel purification using Wizard SV Gel and PCR Clean-Up System (Promega)  
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After worth run an analytic 1% agarose gel and purificate the product with Wizard SV Gel and PCR Clean-Up System (Promega)  
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Revision as of 16:43, 4 October 2013





PCR

Short report
Materials

Equipment
Micropipettes with sterile tips
PCR machine
PCR tubes

Chemicals & consumables
DNA template
Nuclease-free water
Primer
Polymerase buffer
dNTP mix
DMSO

Procedure
This is a commonly used protocol thta we used for Taq-polymerase
PCR program (30 cycles)
initial denaturation 95°C, 60s denaturation 95°C, 45s annealing 54,5°C, 90s elongation 72°C, 120s final elongation 72°C, 120s After worth run an analytic 1% agarose gel and purificate the product with Wizard SV Gel and PCR Clean-Up System (Promega)