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A particular example which demonstrates apparent inconsistencies between the expression estimates obtained from different chip-generations will be given below.
(B) Scatter plot with linear square fitted line showing high correlation between the expression estimates from the two methods (average across replicates), for 1094 TFs across individual representative cell-lines.
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We used edgeR to identify differentially expressed genes between healthy and sick samples from the expression estimates computed with each mapping method.
Table 1 (left) shows the correlation between the log expression estimates of TP transcripts and the true log expression values of the TP transcripts for transcript sets obtained using annotations that have a range of sensitivities and precisions.
For each method, we compute the ratio between the mean expression estimate of FP transcripts and the mean expression estimate of TP transcripts (FP ¯ / TP ¯ ).
There was a strong inverse correlation between the expression levels estimated by mgMOS and the ΔCT value from the RT-PCR (Fig. 3; n = 61, R2 = 0.7, p<0.0001).
We first determined whether we could reproduce the agreement between mRNA expression estimates measured by microarrays and by RNA-Seq reported in other studies.
Examining the dendrogram in Panel C of Figure 3, we observe TopHat2 and Bowtie2 have the shortest distance between gene expression estimates, followed by GSNAP, Bowtie2-G, and Stampy-G, and finally Stampy.
Together with the fact that housekeeping genes had higher expression levels (median 8.29; mean 8.52) than non-housekeeping genes (median 7.46; mean 7.98), the results suggested a possible link between the expression levels and estimates of gene expression heritabilities.
The actin-normalized expression ratios from qRT-PCR were then compared with the actin-normalized expression ratios from RNA-Seq to determine the correlation of actin-normalized expression estimates between the two platforms (supplementary fig. S2, Supplementary Material online).
The association between copy number levels and expression estimates was quantified by fitting a linear model between continuous copy number estimates (segment means) and log2-transformed normalized expression estimates and testing the slopes for significance.
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