Specificity validation of RT-qPCR
Amplifications were successful for all candidate genes in both rat and human specimens
(FigS2.A). Amplification curves of a seven log10 dilution series in human qPCR assays were
displayed in FigS2.B as an example and corresponding standard curve was represented in
FigS2.C.The slope, error and R2 of the regression line and the PCR-efficiency of all candidate
genes were reviewed in Table S2.
Fig S2.A Amplification plot (left) and dissociation curve (right) for the RPS29 assay of
human lung tissues as an example
Fig S2.B Amplification curves of a seven log10 dilution series of RPS29 cDNA templates in
the human qPCR assays.
Fig S2.C Extrapolation of Ct values for RPS29 as function of the log10 of the number of
templates was a straight line (R2= 0.9997) with slope of -3.804 over seven log10 dilution of
the template.
Table S2. A PCR efficiency, slope, error and R2 of regression line of all candidate genes
Gene
GAPDH
β-actin
18SrRNA
RPS29
U6snRNA
5SrRNA
let-7a
MiR-9
MiR-125
Slope
-3.615
-3.558
-3.234
-3.391
-3.425
-3.350
-3.541
-3.246
-3.247
Error
0.0345
0.0321
0.0543
0.0206
0.0085
0.0148
0.0413
0.0348
0.0548
R2
0.9994
0.9995
0.9986
0.9998
0.9999
0.9999
0.9992
0.9994
0.9991
PCR Efficiency
1.891 / 89.1%
1.910 / 91.0%
2.038 / 103.8%
1.972 / 97.2%
1.959 / 95.9%
1.988 / 98.8%
1.916 / 91.6%
2.033 / 103.3%
2.032 / 103.2%