Comments (7)
I see, and in the denominator you used the sum of genes and TEs FPKM?
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Also, it would be helpful if you could clarify that each column means in *_TEcounts.txt file.
i.e. if "fpkm" computed using "uniq_counts" , "tot_counts" or "tot_reads"?
from squire.
Hello,
Sorry for the long response time. Thanks for your feedback; we'll work on building documentation of the output files.
For *_refGenecounts.txt, here are the headers:
chr: chromosome of transcription
tx_start: coordinate for start of transcription
tx_stop: coordinate for end of transcription
Gene_ID: gene name
fpkm: fragments per kilobase per million reads
strand: + or - for stranded data
count: read count, computed by transcript length * coverage / readlength
transcript_ID: transcript specifier
We used the base StringTie parameters for gene expression counting (-M .95) for guided assembly.
We're working on making the full *_TEcounts.txt format documentation, but for the time being, fpkm is computed using total counts.
Thanks for your patience as we sort it out! Let us know if you have other questions in the meantime.
Regards,
Chloe
from squire.
Hi there!
Has this been sorted out? I'm running some analysis on these files, and need a bit of clarification in the *_TEcounts.txt headers.
from squire.
Hello,
I am interested in getting the TPM (transcripts per kilobase million) for TE, which is lacking in the version I am using. I am trying to calculate the TPM from fpkm. (https://haroldpimentel.wordpress.com/2014/05/08/what-the-fpkm-a-review-rna-seq-expression-units/?fbclid=IwAR0gVbmAYOZzIKWtQJU8SHUbQ5Nhh4NaSIi9ObOr0Shoxplft1ZjAv8qPDg)
But before doing so, I want to make sure the FPKM was calculated from the total count of TE and refGene, right?
Many thanks for developing this awesome tool!
from squire.
Hi @MaxwellShih,
I started using SQuiRE recently and I am facing all the problems mentioned in the issues which haven't been corrected yet.
I also would like to use TPM and I thin you should use the read_count field as input for the TPM calculation.
I think you can consider the read_count as the raw_counts.
from squire.
Hello @emattei ,
Thanks for your comments. To calculate TPMs from raw counts, I have to get the gene length information. However, I found it's not easy to get the gene length information, at least for me. So, I just did it from FPKM.
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Related Issues (20)
- Normalization HOT 1
- Generation of final BAM with re-distribute reads according to EM
- Error while running call
- Unable to install squire, package dependency issue HOT 3
- issue with squire count:local variable 'prev_TE_ID' referenced before assignment
- Error with squire Call
- I can´t to install SQuiRE in conda HOT 2
- squire requires two conflicting python versions? HOT 1
- Count does not consider strangeness HOT 1
- Call Output
- T2T Reference Genome HOT 1
- FPKM counts are more than Tot_counts in Squire Count output.
- support Nanopore direct RNA sequencing data ?
- Antisense transcription quantification
- Unable to Install SQuIRE HOT 4
- Using SQuIRE with non-model organism HOT 1
- Expectation Maximization on TE counting
- Sorting mistake?
- bioconductor-deseq2 1.16.1 but there are no viable options
- ATACseq
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