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. 2014 Dec 15;7(12):5947–5957.

Figure 4.

Figure 4

Standard curve in the absolute method of RT-PCR quantification of miRNA. We used standard curve of real-time PCR (RT-PCR) to perform the absolute quantification analysis. Data analysis showed a skewed distribution and the reference values (p=0.05) were determined using percentile method. The ranges of the reference values (copy number /μg total RNA) were as follows: miR-150: 1.5×105~6.9×106; miR-23a: 2.7×108~1.8×109; miR-130a: 2.2×105~1.8×106. In an absolute method, the accuracy was assured by the consistent sample loading (consistent U6 snRNA copy number). From these results, the ranges of ratio of miRNA copy number to U6snRNA copy number in normal human subject blood were also derived: miR-150: 4.71~115.88×10-6; miR-23a: 9.29~56.78×10-3; miR-130a: 7.01~45.72×10-6. Below the normal reference ratio range lower limit would be considered as a reference diagnostic criteria for GC.