I am working with Illumina RNAseq data. My experiment contain 5-5 biological replicates of normal and treated. Initially I followed cufflink-->cuffdiff workflow (Trimmomatics-->Tophat-->Cufflinks-->Cuffmerge-->Cuffdiff), but could not get any significant genes/isoform.
Later I started working working with HTSeq-->EdgeR workflow (Trimmomatics-->Tophat-->HTSeq-->EdgeR) and could get around 1000 significant genes.
Now I am looking for isoform information and so I started working DEXSeq package.
I am getting following error while running dexseq_count.py:
Traceback (most recent call last):
File "/data/RNASEQdata/Programs/dexseq_count.py", line 70, in <module>
for f in HTSeq.GFF_Reader( gff_file ):
File "/home/gadgil/.local/lib/python2.7/site-packages/HTSeq/__init__.py", line 210, in __iter__
for line in FileOrSequence.__iter__( self ):
File "/home/gadgil/.local/lib/python2.7/site-packages/HTSeq/__init__.py", line 51, in __iter__
lines = open( self.fos )
IOError: [Errno 2] No such file or directory: '-'
These are the steps I have followed:
python dexseq_prepare_annotation.py Homo_sapiens.GRCh37.71.gtf DEXSeq.gtf
samtools view accepted_hits.bam | python /dexseq_count.py -p no -s no -a 10 - DEXSeq.gtf output/19_N.txt
Please help me with this error.
Thanks in advance.
Best Regards,
Mudita
Later I started working working with HTSeq-->EdgeR workflow (Trimmomatics-->Tophat-->HTSeq-->EdgeR) and could get around 1000 significant genes.
Now I am looking for isoform information and so I started working DEXSeq package.
I am getting following error while running dexseq_count.py:
Traceback (most recent call last):
File "/data/RNASEQdata/Programs/dexseq_count.py", line 70, in <module>
for f in HTSeq.GFF_Reader( gff_file ):
File "/home/gadgil/.local/lib/python2.7/site-packages/HTSeq/__init__.py", line 210, in __iter__
for line in FileOrSequence.__iter__( self ):
File "/home/gadgil/.local/lib/python2.7/site-packages/HTSeq/__init__.py", line 51, in __iter__
lines = open( self.fos )
IOError: [Errno 2] No such file or directory: '-'
These are the steps I have followed:
python dexseq_prepare_annotation.py Homo_sapiens.GRCh37.71.gtf DEXSeq.gtf
samtools view accepted_hits.bam | python /dexseq_count.py -p no -s no -a 10 - DEXSeq.gtf output/19_N.txt
Please help me with this error.
Thanks in advance.
Best Regards,
Mudita
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