Matt:LabNotes/qPCR Protocol

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qPCR with DNA standards[edit]

DNA Standards

  1. 10 ng/ul
  2. 1 ng/ul
  3. 0.1 ng/ul
  4. 0.01 ng/ul

Procedure: Standard qPCR

  • 1 ul DNA template
  • 15 ul 2x KAPA MasterMix
  • 0.6 ul Forward primer
  • 0.6 ul Reverse primer
  • 12.8 ul H2O

Cycles of: 95C 0:10 58C 0:30 72C 2:00

Removed 2 tubes from each of the 4 samples every time one of their curves began to increase, suggesting that it reached levels differentiable from background noise. The DNA concentrations were measured