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Table 3 Summary of the fluorescence ratio based method and the Ct base method for quantification of predefined standards

From: Quantify single nucleotide polymorphism (SNP) ratio in pooled DNA based on normalized fluorescence real-time PCR

Group No. In predefined standards Predefined allele ratio YMDD: Kf method Baseline subtracted fluorescence ratio SDM
  YIDD Mean (n = 4)a S.D (n = 4)b Intra C.Vc Mean (n = 4)d S.D (n = 4)e Intra C.Vc Mean (n = 4)f S.D (n = 4)g Intra C.Vc
1 90%: 10% 2.68 0.32 11.80% 8.31 0.99 11.95% 6.46 3.06 47.29%
2 80%: 20% 1.40 0.08 5.45% 4.25 0.27 6.45% 2.73 0.09 3.30%
3 70%: 30% 0.80 0.14 17.22% 2.15 0.69 31.87% 1.46 0.14 9.32%
4 60%: 40% 0.54 0.03 6.11% 1.38 0.17 12.22% 1.09 0.27 24.43%
5 50%: 50% 0.40 0.03 7.30% 1.12 0.17 15.59% 0.65 0.07 10.38%
6 40%: 60% 0.30 0.02 7.77% 0.76 0.10 13.09% 0.51 0.04 8.56%
7 30%: 70% 0.22 0.00 1.91% 0.62 0.03 4.89% 0.32 0.08 25.12%
8 20%: 80% 0.15 0.01 3.67% 0.39 0.03 6.57% 0.43 0.18 42.29%
9 10%: 90% 0.07 0.01 10.24% 0.17 0.01 5.98% 0.01 0.02 113.30%
Av. Average CV    7.94%    12.07%    31.56%
  1. The spreadsheet used for this analysis is available as Supplementary Material.1 [see Additional file 1]
  2. a. Arithmetic mean Kf of 4 replicates
  3. b. Combined Standard deviation Kf for 4 replicates
  4. c. (SD/mean) × 100%
  5. d. Fluorescence ratio arithmetic mean of 4 replicates
  6. e. Combined Standard deviation fluorescence ratio for 4 replicates
  7. f. Arithmetic mean 2-ΔCt of 4 replicates
  8. g. Standard deviation of 2-ΔCt for 4 replicates