Haynes Lab:Notebook/SynBERC Scholars 2013/2014/02/14

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Procedure Digital PCR


  • 16 PCR Reactions were run today, following the procedure listed below.
  • TNF A forward and reverse primers were diluted from 100uM to 10uM. Initial stock DNA (Concentration = 2.3mg/mL) was diluted several times to obtain 4 different templates, listed below:
  • D0 = U20S 1:100 = 6.24ng/mL
  • D1 = 0.624ng/mL
  • D2 = 0.0624ng/mL = 6.24*10^(-2) ng/mL
  • D3 = 6.24pg/mL

  • For each PCR reaction, the following volumes will be added:
  • 1uL of forward TNF A primer
  • 1uL of reverse TNF A primer
  • 1uL of diluted DNA (either U20S 1:100, D1, D2, or D3 for each sample)
  • 50uL of PCR master mix
  • 47 uL of dH20
  • 8 samples of each template will be put through the PCR sequence (NOTE: The program to be run is named "First Trial 121713")
  • 2 samples of plain water (instead of template) will also be tested as a negative control.

PCRs Run Today

  • 8 D0 wells
  • 8 D1 wells
  • Labeled and stored in freezer.

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