Figure 1.
Workflow of the study design.
Table 1.
TaqMan probes for the 21 candidate genes analyzed by qRT-PCR.
Table 2.
RNA purity, yield, and integrity.
Figure 2.
qRT-PCR analysis of 19 target transcripts.
GAPDH and the Ctrl1 condition were used as a reference transcript and condition, respectively. ΔΔCt values are shown by vertical axes. Horizontal axes represent pre-analytical conditions in the following order: Ctrl2, Without stab, Protect, Lock, Stab, SDS, and 1-Thio. *, p<0.05; **, p<0.01 (Wilcoxon signed rank test).
Figure 3.
Quality assessment and control of sequencing data from HiSeq2500.
A. Quality values of sequence reads calculated by Cufflinks (Cuffdiff) in each sample. B. The blue bars show the number of sequence reads mapped to the human genome (hs37d5) with TopHat, and the red line with squares indicates the mapped percentage in each sample (B).
Figure 4.
Data bias of transcriptome analysis in each condition.
A. Correlation analysis of the average of FPKM under eight conditions for each sample. B. Cluster analysis of 56 transcriptomes: eight conditions for each of seven volunteers. C. Pair-wise comparisons of significant differences in gene expression for each sample. The number in each box shows the number of differentially expressed genes (p<0.05, Wilcoxon signed rank test).
Figure 5.
Protocol to reduce systematic bias for transcriptome analysis of PBMCs collected in remote assessment centers.
A protocol for pre-analytical operations to mediate the effects of systematic bias in transcriptome data of PBMCs for transportation and biobanking is shown.