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The highly assortative nature of the Always Correlated landscape is clearly evident when node size is mapped to connectivity instead of transcript abundance (Figure 3B).
Thus, we used the coding sequence length instead of transcript length in our calculation: (11) R i b o s o m e d e n s i t y g e n e i = r i b o s o m e n u m b e r o f t h e P e a k F r a c t i o n g e n e i C o d i n g S e q u e n c e l e n g t h g e n e i Ribosome density and ribosome occupancy values are available in Additional file 4: Table S1.
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Such an exon-, instead of transcript-based analysis would offer the advantage of less transcript annotation changes.
In the present study all analyses will be based on genes instead of transcripts, thereby avoiding the bias introduced by different genes having different numbers of transcripts.
By mapping reads to coding sequences instead of transcripts, we tried to increase the accuracy of read assignment to putative genes but lost the information from reads that represent untranslated regions (UTRs).
Instead of capturing transcript molecules by molecular hybridisation, as on microarrays, RNA-seq directly sequences the transcripts present in a sample.
Hence, the increased expression level observed from the microarray data was probably due to the augmented leukocyte number instead of the transcript expression level in the cells.
This approach is similar to the original RPKM definition [ 22] instead of the transcript isoform level estimate as in Cufflinks [ 21].
In the following paper, we will use term feature, instead of probeset, transcript, gene, or protein, to emphasize that the methodology presented has general applicability.
Instead of assembling transcripts separately for each sample and merging them together, our framework reconstructs transcripts directly from multiple samples.
Control plants were mock-inoculated in the same way with water instead of viral transcripts.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com