[2019-04-24 14:49:30] Beginning TopHat run (v2.1.1) ----------------------------------------------- [2019-04-24 14:49:30] Checking for Bowtie Bowtie version: 2.2.4.0 [2019-04-24 14:49:30] Checking for Bowtie index files (genome).. [2019-04-24 14:49:30] Checking for reference FASTA file [2019-04-24 14:49:30] Generating SAM header for Bowtie2Index/genome [2019-04-24 14:49:35] Reading known junctions from GTF file [2019-04-24 14:49:40] Preparing reads left reads: min. length=32, max. length=36, 21100610 kept reads (25510 discarded) right reads: min. length=32, max. length=36, 20963743 kept reads (162377 discarded) [2019-04-24 14:57:22] Building transcriptome data files /scratch/5555363.1.linga/tophat2/tmp/RefSeq_GeneBody [2019-04-24 14:57:40] Building Bowtie index from RefSeq_GeneBody.fa [2019-04-24 15:06:07] Mapping left_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-24 15:20:37] Mapping right_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-24 15:34:57] 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 15:34:58] Mapping left_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-24 15:37:37] Mapping right_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-24 15:41:18] Searching for junctions via segment mapping [2019-04-24 15:43:21] Retrieving sequences for splices [2019-04-24 15:45:18] Indexing splices [2019-04-24 15:45:40] 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/5555363.1.linga/tophat2/tmp/left_kept_reads.m2g_um_unmapped.bam [2019-04-24 15:45:43] Joining segment hits [2019-04-24 15:50:30] 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/5555363.1.linga/tophat2/tmp/right_kept_reads.m2g_um_unmapped.bam [2019-04-24 15:50:35] Joining segment hits [2019-04-24 15:55:01] Reporting output tracks ----------------------------------------------- [2019-04-24 16:17:42] A summary of the alignment counts can be found in /scratch/5555363.1.linga/tophat2/align_summary.txt [2019-04-24 16:17:42] Run complete: 01:28:12 elapsed