Exact(7)
We observe a characteristic coverage bias towards high abundance transcripts in amplification-based sequencing.
More importantly, these reads tended to be less concentrated at the very highest abundance transcripts as shown in Figure 1B, where fraction of total reads mapping to the highest abundance transcripts in SMS are 4% below that of AS.
HXK is presumably encoded by low abundance transcripts in developing guar seeds.
Many of these transcripts may have been previously mistakenly identified as tissue-specific based on EST data only because EST sequencing was not deep enough to detect these low abundance transcripts in different tissues.
These effects may be due to several possible factors, including the propensity of short read lengths to hinder assembly, and incomplete incidence of genes corresponding to low abundance transcripts in current sequence databases [ 20].
This quantitative agreement between theoretical expectation and the data provides clear evidence that sequencing error results in a proliferation of erroneous sequences of high abundance transcripts in next generation sequencing datasets.
Similar(53)
Dscam1 is a low to moderate abundance transcript in embryos (See Author response image 3).
Brain, testis, lung and breast are enriched for genes with abundant transcripts and a shortened 3′ UTR, while low-abundance transcripts in several other tissues (e.g. heart, skeletal muscle) tend to have a lengthened 3′ UTR.
Therefore, we measured HMMSplicer's capacity for detection of junctions in low-abundance transcripts in the human resting CD4 cell dataset.
In addition, limited representation of low-abundance transcripts in dbEST likely has introduces a bias towards SNPs present in genes which display widespread tissue distribution, or are present in tissue types overrepresented in the database.
NGS is also rendering the transcriptome more accessible, with technologies such as Illumina-based RNA-Seq allowing sampling to depths capable of detecting relatively low-abundance transcripts in specific tissues and environmental conditions.
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