Finally have the answer from The Author !!...Though I guessed some of these facts but was in need for confirmation...These facts answer a few other queries regarding the tool too.
Thank you so much Geo.pertea.
Hello People,
I ran the new tuxedo pipeline for a reference based transcriptome analysis on HumanGenome19. After running HISAT2, I executed StringTie with -A option and got the abundance file and it looks like the following
Gene ID Gene Name Reference Strand Start End Coverage FPKM TPM
ENSG00000223972 DDX11L1 1 + 11869 14412 0.261954 0.056163 0.152539
ENSG00000227232 WASH7P 1 - 14363 29806 23.156389 5.585505 15.170202
ENSG00000243485 MIR1302-10 1 + 29554 31109 0.093882 0.048981 0.133031
Now what I would like to know is how is the "Coverage" value in 7th column calculated and in what units (or is it in percentage) ??
In the manual page, it shows "cov: The average per-base coverage for the transcript or exon" but still the calculation is not clear to me.
The coverage value shown in the output of StringTie (and in other genomics programs) is an average of all per-base coverages across the length of genomic segment (exon) or set of segments (transcript). For a gene (which is what is shown in the -A table), the union of all the exonic segments in the gene is considered for this calculation.
For example, an exon coverage value is calculated like this: all the read alignments intersecting the exon are considered and the coverage values for each base (i.e. the number of the read alignments covering that base) are added up and then divided by the length of that exon.
Additionally in StringTie there are a few other factors influencing that per-base coverage value (i.e. the way the number of alignments covering a base is actually "counted"):
Note that the last factor above, although the most complex, does not influence gene coverage values, because at that level we do not need to worry about the distribution of read alignments between transcripts/exons (as mentioned above, simply considering the segment union of all exons in the gene is enough).
Finally have the answer from The Author !!...Though I guessed some of these facts but was in need for confirmation...These facts answer a few other queries regarding the tool too.
Thank you so much Geo.pertea.
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did you already get the answer from somewhere else?
Actually I have tried to look for the answer elsewhere but to no success ...and I also tried to calculate and arrive at the given results by looking at reads mapped at each bases of the shown genes in IGV and still couldn't regenerate the result values !!! ...