[2019-04-29 16:49:28] Beginning TopHat run (v2.1.1) ----------------------------------------------- [2019-04-29 16:49:28] Checking for Bowtie Bowtie version: 2.2.4.0 [2019-04-29 16:49:29] Checking for Bowtie index files (genome).. [2019-04-29 16:49:29] Checking for reference FASTA file [2019-04-29 16:49:29] Generating SAM header for Bowtie2Index/genome [2019-04-29 16:49:33] Reading known junctions from GTF file [2019-04-29 16:49:37] Preparing reads left reads: min. length=35, max. length=43, 5137121 kept reads (18065 discarded) right reads: min. length=35, max. length=43, 5127889 kept reads (27297 discarded) [2019-04-29 16:51:46] Building transcriptome data files /scratch/5765329.1.linga/tophat2/tmp/RefSeq_GeneBody [2019-04-29 16:52:04] Building Bowtie index from RefSeq_GeneBody.fa [2019-04-29 17:00:00] Mapping left_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 17:02:39] Mapping right_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 17:05:18] 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 17:05:18] Mapping left_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 17:08:32] Mapping right_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 17:12:49] Searching for junctions via segment mapping [2019-04-29 17:15:00] Retrieving sequences for splices [2019-04-29 17:17:08] Indexing splices [2019-04-29 17:17:32] 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/5765329.1.linga/tophat2/tmp/left_kept_reads.m2g_um_unmapped.bam [2019-04-29 17:17:36] Joining segment hits [2019-04-29 17:20:43] 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/5765329.1.linga/tophat2/tmp/right_kept_reads.m2g_um_unmapped.bam [2019-04-29 17:20:47] Joining segment hits [2019-04-29 17:23:48] Reporting output tracks ----------------------------------------------- [2019-04-29 17:31:56] A summary of the alignment counts can be found in /scratch/5765329.1.linga/tophat2/align_summary.txt [2019-04-29 17:31:56] Run complete: 00:42:27 elapsed