IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Gold Bioremediation/2010/09/02

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9.2.10

9/2/10 PCR- gesABC & Gas Vesicles

50 uL reactions

1.gesABC

  • 0.8 uL Polymerase
  • 0.5 uL Buffer
  • 0.5 uL DMSO
  • 1.0 uL dNTP
  • 1.0 uL Primer F
  • 1.0 uL Primer R
  • 1.0 uL Template (from PCR Purification)
  • 40.2 uL H2O

2.gesABC

  • 0.8 uL Polymerase
  • 0.5 uL Buffer
  • 0.5 uL DMSO
  • 1.0 uL dNTP
  • 1.0 uL Primer F
  • 1.0 uL Primer R
  • 1.0 uL Template (Genomic)
  • 40.2 uL H2O

3.gesABC

  • 0.8 uL Polymerase
  • 0.5 uL Buffer
  • 0.5 uL DMSO
  • 1.0 uL dNTP
  • 1.0 uL Primer F
  • 1.0 uL Primer R
  • 1.0 uL Template (Genomic)
  • 40.2 uL H2O

4.gesABC

  • 0.8 uL Polymerase
  • 0.5 uL Buffer
  • 0.5 uL DMSO
  • 1.0 uL dNTP
  • 1.0 uL VF2
  • 1.0 uL VR
  • 1.0 uL Template (~13 ng)
  • 40.2 uL H2O

5.gesABC

  • 0.8 uL Polymerase
  • 0.5 uL Buffer
  • 0.5 uL DMSO
  • 1.0 uL dNTP
  • 1.0 uL VR
  • 1.0 uL VF2
  • 1.0 uL Template
  • 40.2 uL H2O

6.gesABC

  • 0.8 uL Polymerase
  • 0.5 uL Buffer
  • 0.5 uL DMSO
  • 1.0 uL dNTP
  • 1.0 uL VF2
  • 1.0 uL VR
  • 41.2 uL H2O

Program:

  • 95 °C - 10 min.
  • 95 °C – 30 sec.
  • 56 °C – 30 sec.
  • 72 °C – 6 min.
  • Go to Step 2 – 35 times
  • 72 °C – 15 min.
  • Hold 4 °C




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