[2019-04-24 16:17:09] Beginning TopHat run (v2.1.1) ----------------------------------------------- [2019-04-24 16:17:09] Checking for Bowtie Bowtie version: 2.2.4.0 [2019-04-24 16:17:09] Checking for Bowtie index files (genome).. [2019-04-24 16:17:09] Checking for reference FASTA file [2019-04-24 16:17:09] Generating SAM header for Bowtie2Index/genome [2019-04-24 16:17:13] Reading known junctions from GTF file [2019-04-24 16:17:17] Preparing reads left reads: min. length=32, max. length=36, 18147441 kept reads (115459 discarded) right reads: min. length=32, max. length=36, 17941404 kept reads (321496 discarded) [2019-04-24 16:23:45] Building transcriptome data files /scratch/5555378.1.linga/tophat2/tmp/RefSeq_GeneBody [2019-04-24 16:24:03] Building Bowtie index from RefSeq_GeneBody.fa [2019-04-24 16:31:47] Mapping left_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-24 16:40:14] Mapping right_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-24 16:48:07] 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 [2019-04-24 16:48:07] Mapping left_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-24 16:49:55] Mapping right_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-24 16:54:13] Searching for junctions via segment mapping [2019-04-24 16:56:13] Retrieving sequences for splices [2019-04-24 16:58:18] Indexing splices [2019-04-24 16:58:38] 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/5555378.1.linga/tophat2/tmp/left_kept_reads.m2g_um_unmapped.bam [2019-04-24 16:58:42] Joining segment hits [2019-04-24 17:02:15] 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/5555378.1.linga/tophat2/tmp/right_kept_reads.m2g_um_unmapped.bam [2019-04-24 17:02:19] Joining segment hits [2019-04-24 17:05:49] Reporting output tracks ----------------------------------------------- [2019-04-24 17:20:13] A summary of the alignment counts can be found in /scratch/5555378.1.linga/tophat2/align_summary.txt [2019-04-24 17:20:13] Run complete: 01:03:04 elapsed