[2019-04-29 16:39:16] Beginning TopHat run (v2.1.1) ----------------------------------------------- [2019-04-29 16:39:16] Checking for Bowtie Bowtie version: 2.2.4.0 [2019-04-29 16:39:17] Checking for Bowtie index files (genome).. [2019-04-29 16:39:17] Checking for reference FASTA file [2019-04-29 16:39:17] Generating SAM header for Bowtie2Index/genome [2019-04-29 16:39:19] Reading known junctions from GTF file [2019-04-29 16:39:22] Preparing reads left reads: min. length=35, max. length=43, 5045393 kept reads (18276 discarded) right reads: min. length=35, max. length=43, 5033248 kept reads (30421 discarded) [2019-04-29 16:40:57] Building transcriptome data files /scratch/5765327.1.c/tophat2/tmp/RefSeq_GeneBody [2019-04-29 16:41:07] Building Bowtie index from RefSeq_GeneBody.fa [2019-04-29 16:46:36] Mapping left_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 16:48:16] Mapping right_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-29 16:49:30] 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 16:49:30] Mapping left_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 16:50:17] Mapping right_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-29 16:50:50] Searching for junctions via segment mapping [2019-04-29 16:51:55] Retrieving sequences for splices [2019-04-29 16:53:05] Indexing splices [2019-04-29 16:53:17] 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/5765327.1.c/tophat2/tmp/left_kept_reads.m2g_um_unmapped.bam [2019-04-29 16:53:19] Joining segment hits [2019-04-29 16:54:57] 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/5765327.1.c/tophat2/tmp/right_kept_reads.m2g_um_unmapped.bam [2019-04-29 16:54:59] Joining segment hits [2019-04-29 16:56:32] Reporting output tracks ----------------------------------------------- [2019-04-29 17:00:19] A summary of the alignment counts can be found in /scratch/5765327.1.c/tophat2/align_summary.txt [2019-04-29 17:00:19] Run complete: 00:21:02 elapsed