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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