Hello sachit, Sachit Adhikari <sachit.technerves@...> writes:

I am getting this error:

Error in tophat:

[2013-02-13 20:46:41] Beginning TopHat run (v2.0.7) ----------------------------------------------- [2013-02-13 20:46:41] Checking for Bowtie Bowtie version: 2.0.6.0 [2013-02-13 20:46:41] Checking for Samtools Samtools version: 0.1.18.0 [2013-02-13 20:46:41] Checking for Bowtie index files [2013-02-13 20:46:41] Checking for reference FASTA file Warning: Could not find FASTA file

[2013-02-13 20:46:41] Reconstituting reference FASTA file from Bowtie index Executing: /usr/bin/bowtie2-inspect /data/rathi/Homo_sapiens/UCSC/hg19/Sequence/Bowtie2Index/genome > ./tophat_out/tmp/genome.fa [2013-02-13 20:48:51] Generating SAM header for /data/rathi/Homo_sapiens/UCSC/hg19/Sequence/Bowtie2Index/genome format: fastq quality scale: phred33 (default) [2013-02-13 20:49:23] Preparing reads left reads: min. length=34, max. length=34, 2 kept reads (0 discarded) Warning: you have only one segment per read. If the read length is greater than or equal to 45bp, we strongly recommend that you decrease --segment-length to about half

[2013-02-13 20:49:23] Mapping left_kept_reads to genome genome with Bowtie2 [2013-02-13 20:49:56] Searching for junctions via segment mapping Coverage-search algorithm is turned on, making this step very slow Please try running TopHat again with the option (--no-coverage-search) if

Warning: junction database is empty! [2013-02-13 20:51:18] Reporting output tracks [FAILED] Error running /usr/local/bin/tophat_reports --min-anchor 8 --splice-mismatches 0 --min-report-intron 50 --max-report-intron 500000 --min-isoform-fraction 0.15 --output-dir ./tophat_out/ --max-multihits 20 --max-seg-multihits 40 --segment-length 25 --segment-mismatches 2 --min-closure-exon 100 --min-closure-intron 50 --max-closure-intron 5000 --min-coverage-intron 50 --max-coverage-intron 20000 --min-segment-intron 50 --max-segment-intron 500000 --read-mismatches 2 --read-gap-length 2 --read-edit-dist 2 --read-realign-edit-dist 3 --max-insertion-length 3 --max-deletion-length 3 -z gzip -p4 --no-closure-search --no-microexon-search --sam-header ./tophat_out/tmp/genome_genome.bwt.samheader.sam --report-discordant-pair-alignments --report-mixed-alignments --samtools=/bin/samtools --bowtie2-max-penalty 6 --bowtie2-min-penalty 2 --bowtie2-penalty-for-N 1 --bowtie2-read-gap-open 5 --bowtie2-read-gap-cont 3 --bowtie2-ref-gap-open 5 --bowtie2-ref-gap-cont 3 ./tophat_out/tmp/genome.fa ./tophat_out/junctions.bed ./tophat_out/insertions.bed ./tophat_out/deletions.bed ./tophat_out/fusions.out ./tophat_out/tmp/accepted_hits ./tophat_out/tmp/left_kept_reads.bam Loading ...done

What's wrong?

<div> <p>I am getting this error:</p> <div><br></div> <div>Error in tophat:

[2013-02-13 20:46:41] Beginning TopHat run (v2.0.7) ----------------------------------------------- [2013-02-13 20:46:41] Checking for Bowtie Bowtie version: 2.0.6.0 [2013-02-13 20:46:41] Checking for Samtools Samtools version: 0.1.18.0 [2013-02-13 20:46:41] Checking for Bowtie index files [2013-02-13 20:46:41] Checking for reference FASTA file Warning: Could not find FASTA file /data/rathi/Homo_sapiens/UCSC/hg19/Sequence/Bowtie2Index/genome.fa [2013-02-13 20:46:41] Reconstituting reference FASTA file from Bowtie index Executing: /usr/bin/bowtie2-inspect /data/rathi/Homo_sapiens/UCSC/hg19/Sequence/Bowtie2Index/genome > ./tophat_out/tmp/genome.fa [2013-02-13 20:48:51] Generating SAM header for /data/rathi/Homo_sapiens/UCSC/hg19/Sequence/Bowtie2Index/genome format: fastq quality scale: phred33 (default) [2013-02-13 20:49:23] Preparing reads left reads: min. length=34, max. length=34, 2 kept reads (0 discarded) Warning: you have only one segment per read. If the read length is greater than or equal to 45bp, we strongly recommend that you decrease --segment-length to about half

[2013-02-13 20:49:23] Mapping left_kept_reads to genome genome with Bowtie2 [2013-02-13 20:49:56] Searching for junctions via segment mapping Coverage-search algorithm is turned on, making this step very slow Please try running TopHat again with the option (--no-coverage-search) if

/data/rathi/Homo_sapiens/UCSC/hg19/Sequence/Bowtie2Index/genome.fa the read length because TopHat will work better with multiple segments this step takes too much time or memory. the read length because TopHat will work better with multiple segments this step takes too much time or memory.

Warning: junction database is empty! [2013-02-13 20:51:18] Reporting output tracks [FAILED] Error running /usr/local/bin/tophat_reports --min-anchor 8 --splice-mismatches 0 --min-report-intron 50 --max-report-intron 500000 --min-isoform-fraction 0.15 --output-dir ./tophat_out/ --max-multihits 20 --max-seg-multihits 40 --segment-length 25 --segment-mismatches 2 --min-closure-exon 100 --min-closure-intron 50 --max-closure-intron 5000 --min-coverage-intron 50 --max-coverage-intron 20000 --min-segment-intron 50 --max-segment-intron 500000 --read-mismatches 2 --read-gap-length 2 --read-edit-dist 2 --read-realign-edit-dist 3 --max-insertion-length 3 --max-deletion-length 3 -z gzip -p4 --no-closure-search --no-microexon-search --sam-header ./tophat_out/tmp/genome_genome.bwt.samheader.sam --report-discordant-pair-alignments --report-mixed-alignments --samtools=/bin/samtools --bowtie2-max-penalty 6 --bowtie2-min-penalty 2 --bowtie2-penalty-for-N 1 --bowtie2-read-gap-open 5 --bowtie2-read-gap-cont 3 --bowtie2-ref-gap-open 5 --bowtie2-ref-gap-cont 3 ./tophat_out/tmp/genome.fa ./tophat_out/junctions.bed ./tophat_out/insertions.bed ./tophat_out/deletions.bed ./tophat_out/fusions.out ./tophat_out/tmp/accepted_hits ./tophat_out/tmp/left_kept_reads.bam Loading ...done </div> <div>What's wrong?</div> </div>

even I am getting the same error. please let me know if you have solved it.