[2019-04-29 16:58:36] Beginning TopHat run (v2.1.1) ----------------------------------------------- [2019-04-29 16:58:36] Checking for Bowtie Bowtie version: 2.2.4.0 [2019-04-29 16:58:37] Checking for Bowtie index files (genome).. [2019-04-29 16:58:37] Checking for reference FASTA file [2019-04-29 16:58:37] Generating SAM header for Bowtie2Index/genome [2019-04-29 16:58:41] Reading known junctions from GTF file [2019-04-29 16:58:45] Preparing reads left reads: min. length=35, max. length=43, 5551815 kept reads (22601 discarded) right reads: min. length=35, max. length=43, 5549141 kept reads (25275 discarded) [2019-04-29 17:01:03] Building transcriptome data files /scratch/5765337.1.linga/tophat2/tmp/RefSeq_GeneBody [2019-04-29 17:01:18] Building Bowtie index from RefSeq_GeneBody.fa [2019-04-29 17:08:18] Mapping left_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 17:11:02] Mapping right_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 17:13:54] 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 17:13:54] Mapping left_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 17:15:19] Mapping right_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 17:16:31] Searching for junctions via segment mapping [2019-04-29 17:18:25] Retrieving sequences for splices [2019-04-29 17:20:26] Indexing splices [2019-04-29 17:20:49] 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/5765337.1.linga/tophat2/tmp/left_kept_reads.m2g_um_unmapped.bam [2019-04-29 17:20:53] Joining segment hits [2019-04-29 17:23:59] 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/5765337.1.linga/tophat2/tmp/right_kept_reads.m2g_um_unmapped.bam [2019-04-29 17:24:03] Joining segment hits [2019-04-29 17:27:00] Reporting output tracks ----------------------------------------------- [2019-04-29 17:35:03] A summary of the alignment counts can be found in /scratch/5765337.1.linga/tophat2/align_summary.txt [2019-04-29 17:35:03] Run complete: 00:36:26 elapsed