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Table 2 Comparison of reverse transcription (RT) and pre-amplification (PRE) variability

From: Pre-amplification in the context of high-throughput qPCR gene expression experiment

  RND1 CD83 EIF3M STK10 FKBP AVG var.
Efficiency (E) 91% 100% 97% 94% 100%  
Cq RT 31.7 25.6 24.6 22.1 20.2  
\( S{D}_{\mathrm{RT}}=\sqrt{SD{(rt)}^2+SD{(qPCR)}^2} \) 0.38 0.41 0.24 0.40 0.29  
Variability RT = (1 + E)SD RT 28% 33% 18% 17% 22% 23.6%
Cq PRE* 23.6 17.1 16.2 13.4 12.1  
\( S{D}_{PRE}=\sqrt{SD{(pre)}^2+SD{(qPCR)}^2} \) 0.24 0.18 0.18 0.14 0.16  
Variability PRE = (1 + E)SD PRE 17% 13% 13% 10% 12% 13%
Expression Differential 1.2 0.7 1.0 0.7 1.3  
  1. *Equivalent of 5 ng of total RNA was used in 18 cycle pre-amplification, pre-amplified cDNA diluted 40x.