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Table 3 Normalization of EGFR, HER2 and HER3 expression according to the RIN

From: Reverse transcription-quantitative polymerase chain reaction: description of a RIN-based algorithm for accurate data normalization

Cell line RIN EGFR HER2 HER3
   Measured ratio (% error) RIN-normalized ratio (% error) Measured ratio (% error) RIN-normalized ratio (% error) Measured ratio (% error) RIN-normalized ratio (% error)
LS174T 10.0 1.00 (0.0) - 1.00 (0.0) - 1.00 (0.0) -
  8.0 0.75 (33.3) 0.91 (9.9) 0.76 (31.6) 0.93 (7.5) 0.83 (20.5) 1.01 (1.0)
  7.1 0.63 (58.7) 0.84 (19.0) 0.74 (35.1) 0.99 (1.0) 0.63 (58.7) 0.84 (19.0)
  6.6 0.60 (66.7) 0.85 (1.6) 0.63 (58.7) 0.89 (12.4) 0.62 (61.3) 0.87 (14.9)
  5.4 0.50 (100.0) 0.81 (23.5) 0.60 (66.7) 0.98 (2.0) 0.55 (81.8) 0.89 (12.4)
SKBr3 10.0 1.00 (0.0) - 1.00 (0.0) - 1.00 (0.0) -
  7.9 0.84 (19.0) 1.03 (3.0) 0.80 (25.0) 0.99 (1.0) 0.85 (17.6) 1.05 (5.0)
  7.2 0.73 (37.0) 0.96 (4.2) 0.68 (47.0) 0.90 (11.1) 0.74 (35.1) 0.98 (2.0)
  5.9 0.64 (56.2) 0.98 (2.0) 0.61 (63.9) 0.93 (7.5) 0.72 (38.9) 1.10 (10.0)
  5.1 0.56 (78.6) 0.95 (5.2) 0.57(75.4) 0.96 (4.2) 0.63 (58.7) 1.07 (7.0)
  1. EGFR, HER2 and HER3 gene transcription levels were assessed in LS174T and SKBr3 cell samples with decreasing RNA integrity and expressed as n-fold difference relative to the intact (RIN = 10) sample. RIN-normalized ratios were calculated according to the following formula: RIN-normalized ratio = Measured ratio + (Measured ratio × (1-(0.08 × RIN + 0.19))/(0.08 × RIN + 0.19)). The percent of error (% error) shows the accuracy of the estimated (i.e. measured or normalized) ratio and was calculated as follow: % error = ((expected ratio – estimated ratio)/estimated ratio) × 100%.