Team:TU Darmstadt/protocols/PCR

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<h2><font size="6" color="#F0F8FF" face="Arial regular"> PCR </font></h2>
<h2><font size="6" color="#F0F8FF" face="Arial regular"> PCR </font></h2>
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<B> Short report<br></B>
<B> Short report<br></B>
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PCR (polymerase chain reaction) is a method for exponentially amplifying a fragment of DNA in vitro.
<B> Materials<br></B>
<B> Materials<br></B>
<br>
<br>
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<br>
<br>
<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>
<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>
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initial denaturation 95°C, 60s
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initial denaturation 95°C, 60s<br>
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denaturation 95°C, 45s
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denaturation 95°C, 45s<br>
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annealing 54,5°C, 90s
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annealing 54,5°C, 90s <br>
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elongation 72°C, 120s
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elongation 72°C, 120s<br>
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final elongation 72°C, 120s
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final elongation 72°C, 120s<br>
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<br>
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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)  
<br>
<br>

Latest revision as of 00:37, 5 October 2013







Lab book | Materials | Protocols

PCR

Short report
 PCR (polymerase chain reaction) is a method for exponentially amplifying a fragment of DNA in vitro. 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)

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