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Figure 8 | BMC Genomics

Figure 8

From: An optimized protocol for microarray validation by quantitative PCR using amplified amino allyl labeled RNA

Figure 8

Correlation between microarrays and qPCR. Gene expression profiles of blood cells from 15 brain-dead organ donors were analyzed by microarrays covering 25,000-genes. Among these, the following were considered in these experiments: TNF-α, LY96, ELANE, ANXA1 and STAT3. Expression levels of these 5 genes obtained by microarrays were calculated as the log ratio sample vs universal reference RNA. 100 ng of AA-aRNA remaining from microarray experiments were reverse transcribed using either the original or the optimized RT protocol and assessed for expression levels of these genes by qPCR. GAPDH was used as reference gene for normalization. For each gene, expression data from microarrays (log ratio gene/reference) were plotted against expression data from qPCR calculated with the formula (2Cq gene - Cq GAPDH). Correlation between microarrays and qPCR, either using the original () or the optimized () RT protocol, were calculated by Pearson product moment correlation and are summarized in Table 4. Linear regression lines are displayed for correlations between microarrays and qPCR data obtained with the original (full lines) or the optimized (dotted lines) RT protocol. RT protocol optimization did not modify the correlation between microarrays and qPCR.

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