[2019-04-29 17:13:58] Beginning TopHat run (v2.1.1) ----------------------------------------------- [2019-04-29 17:13:58] Checking for Bowtie Bowtie version: 2.2.4.0 [2019-04-29 17:13:59] Checking for Bowtie index files (genome).. [2019-04-29 17:13:59] Checking for reference FASTA file [2019-04-29 17:13:59] Generating SAM header for Bowtie2Index/genome [2019-04-29 17:15:24] Reading known junctions from GTF file [2019-04-29 17:15:31] Preparing reads left reads: min. length=35, max. length=43, 5542813 kept reads (21533 discarded) right reads: min. length=35, max. length=43, 5529718 kept reads (34628 discarded) [2019-04-29 17:18:18] Building transcriptome data files /scratch/5765344.1.linga/tophat2/tmp/RefSeq_GeneBody [2019-04-29 17:19:00] Building Bowtie index from RefSeq_GeneBody.fa [2019-04-29 17:27:06] Mapping left_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 17:30:24] Mapping right_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 17:34:00] Resuming TopHat pipeline with unmapped reads 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 the read length because TopHat will work better with multiple segments [2019-04-29 17:34:00] Mapping left_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 17:35:51] Mapping right_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 17:37:19] Searching for junctions via segment mapping [2019-04-29 17:39:25] Retrieving sequences for splices [2019-04-29 17:41:27] Indexing splices [2019-04-29 17:41:48] Mapping left_kept_reads.m2g_um_unmapped to genome segment_juncs with Bowtie2 (1/1) open: No such file or directory Error: bam2fastx failed to open BAM file /scratch/5765344.1.linga/tophat2/tmp/left_kept_reads.m2g_um_unmapped.bam [2019-04-29 17:41:52] Joining segment hits [2019-04-29 17:45:20] Mapping right_kept_reads.m2g_um_unmapped to genome segment_juncs with Bowtie2 (1/1) open: No such file or directory Error: bam2fastx failed to open BAM file /scratch/5765344.1.linga/tophat2/tmp/right_kept_reads.m2g_um_unmapped.bam [2019-04-29 17:45:25] Joining segment hits [2019-04-29 17:48:41] Reporting output tracks ----------------------------------------------- [2019-04-29 17:57:06] A summary of the alignment counts can be found in /scratch/5765344.1.linga/tophat2/align_summary.txt [2019-04-29 17:57:06] Run complete: 00:43:07 elapsed