[2019-04-29 16:56:35] Beginning TopHat run (v2.1.1) ----------------------------------------------- [2019-04-29 16:56:35] Checking for Bowtie Bowtie version: 2.2.4.0 [2019-04-29 16:56:36] Checking for Bowtie index files (genome).. [2019-04-29 16:56:36] Checking for reference FASTA file [2019-04-29 16:56:36] Generating SAM header for Bowtie2Index/genome [2019-04-29 16:56:41] Reading known junctions from GTF file [2019-04-29 16:56:45] Preparing reads left reads: min. length=35, max. length=43, 5560415 kept reads (18615 discarded) right reads: min. length=35, max. length=43, 5547640 kept reads (31390 discarded) [2019-04-29 16:59:04] Building transcriptome data files /scratch/5765334.1.linga/tophat2/tmp/RefSeq_GeneBody [2019-04-29 16:59:21] Building Bowtie index from RefSeq_GeneBody.fa [2019-04-29 17:07:49] Mapping left_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 17:10:43] Mapping right_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 17:14:30] 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 17:14:30] Mapping left_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 17:17:02] Mapping right_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 17:18:13] Searching for junctions via segment mapping [2019-04-29 17:20:23] Retrieving sequences for splices [2019-04-29 17:22:19] Indexing splices [2019-04-29 17:22:39] 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/5765334.1.linga/tophat2/tmp/left_kept_reads.m2g_um_unmapped.bam [2019-04-29 17:22:43] Joining segment hits [2019-04-29 17:25:58] 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/5765334.1.linga/tophat2/tmp/right_kept_reads.m2g_um_unmapped.bam [2019-04-29 17:26:08] Joining segment hits [2019-04-29 17:29:57] Reporting output tracks ----------------------------------------------- [2019-04-29 17:38:58] A summary of the alignment counts can be found in /scratch/5765334.1.linga/tophat2/align_summary.txt [2019-04-29 17:38:58] Run complete: 00:42:22 elapsed