[2019-04-29 17:07:44] Beginning TopHat run (v2.1.1) ----------------------------------------------- [2019-04-29 17:07:44] Checking for Bowtie Bowtie version: 2.2.4.0 [2019-04-29 17:07:45] Checking for Bowtie index files (genome).. [2019-04-29 17:07:45] Checking for reference FASTA file [2019-04-29 17:07:45] Generating SAM header for Bowtie2Index/genome [2019-04-29 17:07:51] Reading known junctions from GTF file [2019-04-29 17:08:10] Preparing reads left reads: min. length=35, max. length=43, 5125031 kept reads (22898 discarded) right reads: min. length=35, max. length=43, 5120917 kept reads (27012 discarded) [2019-04-29 17:09:50] Building transcriptome data files /scratch/5765339.1.p8/tophat2/tmp/RefSeq_GeneBody [2019-04-29 17:09:59] Building Bowtie index from RefSeq_GeneBody.fa [2019-04-29 17:14:55] Mapping left_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 17:16:20] Mapping right_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 17:17:30] 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 17:17:31] Mapping left_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 17:18:12] Mapping right_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 17:18:45] Searching for junctions via segment mapping [2019-04-29 17:19:43] Retrieving sequences for splices [2019-04-29 17:20:43] Indexing splices [2019-04-29 17:20:54] 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/5765339.1.p8/tophat2/tmp/left_kept_reads.m2g_um_unmapped.bam [2019-04-29 17:20:56] Joining segment hits [2019-04-29 17:22:27] 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/5765339.1.p8/tophat2/tmp/right_kept_reads.m2g_um_unmapped.bam [2019-04-29 17:22:28] Joining segment hits [2019-04-29 17:23:55] Reporting output tracks ----------------------------------------------- [2019-04-29 17:27:27] A summary of the alignment counts can be found in /scratch/5765339.1.p8/tophat2/align_summary.txt [2019-04-29 17:27:27] Run complete: 00:19:42 elapsed