Team:WashU/Protocols/PCR

From 2012.igem.org

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==Set up PCR reaction tube==
==Set up PCR reaction tube==
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##1uL Template
+
#1uL Template
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##1uL 25mM dNTP's
+
#1uL 25mM dNTP's
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##10uL Buffer
+
#10uL Buffer
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##0.5uL Forward Primer
+
#0.5uL Forward Primer
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##0.5uL Reverse Primer
+
#0.5uL Reverse Primer
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##0.5uL TAQ polymerase
+
#0.5uL TAQ polymerase
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##36.5uL diH2O
+
#36.5uL diH2O
==Amplification reaction in thermocycler==
==Amplification reaction in thermocycler==
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##1 cycle 94 degrees (3 minutes)
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#1 cycle 94 degrees (3 minutes)
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##40 cycles 94 degrees (1 minute), 50 degrees (1 minute), 72 degrees (1 minute)
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#40 cycles 94 degrees (1 minute), 50 degrees (1 minute), 72 degrees (1 minute)
==Source==
==Source==
Thanks to UW iGEM '11 for the above instructions.  The original source can be found here: [https://2011.igem.org/Team:Washington/Protocols/gel_electrophoresis University Washington gel electrophoresis protocol]
Thanks to UW iGEM '11 for the above instructions.  The original source can be found here: [https://2011.igem.org/Team:Washington/Protocols/gel_electrophoresis University Washington gel electrophoresis protocol]

Revision as of 17:12, 18 June 2012


Contents

General PCR Protocol

Set up PCR reaction tube

  1. 1uL Template
  2. 1uL 25mM dNTP's
  3. 10uL Buffer
  4. 0.5uL Forward Primer
  5. 0.5uL Reverse Primer
  6. 0.5uL TAQ polymerase
  7. 36.5uL diH2O

Amplification reaction in thermocycler

  1. 1 cycle 94 degrees (3 minutes)
  2. 40 cycles 94 degrees (1 minute), 50 degrees (1 minute), 72 degrees (1 minute)

Source

Thanks to UW iGEM '11 for the above instructions. The original source can be found here: University Washington gel electrophoresis protocol