Entering edit mode
4.5 years ago
mar77
▴
40
I'm trying to run IsoEM2 for the quantification of my RNA-seq experiment using the command:
isoem2 [--GTF Homo_sapiens.GRCh38.99.chr.gtf]* [--auto-fragment-distrib]* SAMPLE.sam
as per their manual, but then the output I get is just:
isoem v 2.0.0
Usage: isoem [global options]* [library options]* <aligned_reads.sam>
Or: at <aligned_reads.sam> | isoem2 [global options]* [library options]*
Mandatory global options:
------------------------
-G, --GTF <GTF file> Known genes and isoforms in GTF format
Mandatory library options: either -a or both -m and -d must be present:
-------------------------
-m, --fragment-mean <Double> Fragment length mean
-d, --fragment-std-dev <Double> Fragment length standard deviation
-a, --auto-fragment-distrib Automatically detect fragment length
distribution from uniquely mapping
paired reads (DOES NOT WORK FOR
SINGLE READS)
Optional global options:
-----------------------
-c, --gene-clusters <Cluster file> Override isoform to gene mapping
defined in the GTF file with a
mapping taken from the given file.
The format of each line in the file
is "isoform gene"
-g <genome fasta file> Genome reference sequence (needed by
some library options)
-b Perform hexamer bias correction
-h, --help Show help
-r <Repeats GTF> Drop alignments falling within
annotated repeats
Optional library options:
------------------------
-s, --directional Dataset obtained by directed RNA-Seq
(the strand of each read is
deterministically chosen: for single
reads, the read always comes from
the coding strand; for paired reads,
the first read always comes from the
coding strand, the second from the
opposite strand)
--antisense Directional sequencing but the reads
come from the antisense
--mate-pairs Paired reads come from the same strand
(as opposed to the default behavior
where the two reads in a pair are
assumed to come from opposite
strands)
--max-mismatches <Integer> Maximum number of mismatched allowed
for a read. This requires the genome
sequence to be specified (see -g).
-q, --quality-scores Weigh the reads based on their quality
scores. This requires the genome
sequence to be specified (see -g).
--repeat-threshold <nbases> Drop all reads that have more than
this many bases inside annotated
repeats. Default: 20.
--polyA <nbases> Reads have been generated from mRNAs
with polyA tails of approximately
the given number of bases
-o <file prefix> Output files prefix. It can include
path. Default: same as sam file name
-O <directory prefix> Output directory prefix. If read
alignments are read from stdin,
the default value is stdinSample
-C <confidence interval (%)> Compute expression of genes/isoforms
with specified confidence intervals.
Provide an integer (default: 95,
bootstraps: 200)
--endseq Disable length normalization for data
generated using 5' or 3' end-sequen-
cing protocols, which generate a
single fragment per cDNA molecule
It basically just prints out the manual, which does say to use isoem rather than isoem2 as the command, but when I have tried that it says
isoem: command not found
Does anyone have any idea where I might be going wrong and how to fix it?
Is this the literal command you are running:
Or are you running: