rsem-prepare-reference -p 5 --bowtie2 cd_hit_est.fasta ref_output_12
And the files i obtained are:
ref_12.1.bt2
ref_12.2.bt2
ref_12.3.bt2
ref_12.4.bt2
ref_12.grp
ref_12.idx.fa
ref_12.n2g.idx.fa
ref_12.rev.1.bt2
ref_12.rev.2.bt2
ref_12.seq
ref_12.ti
ref_12.transcripts.fa
then i tried running the command:
rsem-calculate-expression -p 5 --bowtie2 --ci-memory 5000 --paired-end SRR5183402_1.fastq SRR5183402_2.fastq ref_output_12 SSE_fm
and this is the error I am getting:
bowtie2 -q --phred33 --sensitive --dpad 0 --gbar 99999999 --mp 1,1 --np 1 --score-min L,0,-0.1 -I 1 -X 1000 --no-mixed --no-discordant -p 5 -k 200 -x ref_output_12 -1 SRR5183402_1.fastq -2 SRR5183402_2.fastq 2> SSE_fm.log | samtools view -b -o SSE_fm.temp/SSE_fm.bam -
[main_samview] fail to read the header from "-".
"bowtie2 -q --phred33 --sensitive --dpad 0 --gbar 99999999 --mp 1,1 --np 1 --score-min L,0,-0.1 -I 1 -X 1000 --no-mixed --no-discordant -p 5 -k 200 -x ref_output_12 -1 SRR5183402_1.fastq -2 SRR5183402_2.fastq 2> SSE_fm.log | samtools view -b -o SSE_fm.temp/SSE_fm.bam -" failed! Plase check if you provide correct parameters/options for the pipeline!
Previosly I had used trinity output, but i got same error. This time i am using clustered data, still the same error.
I have used bowtie2 option while creating both reference and rsem-calculate-expression.
Any help on this would be highly appreciated. Thank you very much.
Best regards,
Bhoomika