[2019-04-29 17:21:35] Beginning TopHat run (v2.1.1) ----------------------------------------------- [2019-04-29 17:21:35] Checking for Bowtie Bowtie version: 2.2.4.0 [2019-04-29 17:21:36] Checking for Bowtie index files (genome).. [2019-04-29 17:21:36] Checking for reference FASTA file [2019-04-29 17:21:36] Generating SAM header for Bowtie2Index/genome [2019-04-29 17:21:40] Reading known junctions from GTF file [2019-04-29 17:21:44] Preparing reads left reads: min. length=35, max. length=43, 6169781 kept reads (35774 discarded) right reads: min. length=35, max. length=43, 6182711 kept reads (22844 discarded) [2019-04-29 17:24:39] Building transcriptome data files /scratch/5765356.1.linga/tophat2/tmp/RefSeq_GeneBody [2019-04-29 17:24:58] Building Bowtie index from RefSeq_GeneBody.fa [2019-04-29 17:33:21] Mapping left_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 17:37:07] Mapping right_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 17:41:13] 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 17:41:13] Mapping left_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 17:42:49] Mapping right_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 17:44:16] Searching for junctions via segment mapping [2019-04-29 17:46:21] Retrieving sequences for splices [2019-04-29 17:48:32] Indexing splices [2019-04-29 17:48:56] 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/5765356.1.linga/tophat2/tmp/left_kept_reads.m2g_um_unmapped.bam [2019-04-29 17:48:59] Joining segment hits [2019-04-29 17:52:26] 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/5765356.1.linga/tophat2/tmp/right_kept_reads.m2g_um_unmapped.bam [2019-04-29 17:52:30] Joining segment hits [2019-04-29 17:55:44] Reporting output tracks ----------------------------------------------- [2019-04-29 18:05:23] A summary of the alignment counts can be found in /scratch/5765356.1.linga/tophat2/align_summary.txt [2019-04-29 18:05:23] Run complete: 00:43:48 elapsed