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We performed reverse transcription coupled with qPCR assays on a subset of genes to verify the differential expression estimated with microarray analysis and to determine the degree of change in transcript abundance of those genes in individual samples in two subsequent seasons.
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However, their directions of expression change estimated by qRT-PCR are consistent with those estimated from microarray (Figure 6).
Even though MSN and TBX2 genes have less than a 1.5-fold change in expression level based on qRT-PCR in USC vs. EAC-G1 comparison, the direction (down or up-regulated) of their expression estimated by qRT-PCR are consistent with those estimated from microarray (Fig 5c d).
Similar results were obtained with the 636 immune-related transcripts estimated by microarray hybridization.
Given the network topology, p can be estimated from microarray experiments by standard Bayessian parametric inference with two states (see Methods).
We have shown that sequencing-based estimates of transcript abundance are in good agreement with estimates acquired with microarrays, provided that sequencing depth is sufficient [ 48].
Most G+C content variables show much stronger correlations with the RPKM values than are with the expression levels estimated from the microarray data.
Since the most significant differences between Figures 1 and 2 occurs in the range of 0 to 200 ppm, we calculated the correlation coefficient Eq. 2 only for those genes with microarray estimated expression rates less than 200 ppm.
Table 1 compares sensitivity and specificity figures estimated using genotype microarray data (fourth column) and our method with two different sets of allele frequencies (second and third column).
A simple linear regression analysis was performed to correlate fold changes calculated by qRT-PCR analysis with fold changes estimated from the microarrays using the statistical computing.
The concordance of our RNA-seq based estimates with previous microarray-based data from other bird species is in contrast with findings from Xiong et al. 2010 [ 22] who report a striking difference between microarray and RNA-seq based estimates of X- to autosomal expression ratios.
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