[2019-04-24 14:47:48] Beginning TopHat run (v2.1.1) ----------------------------------------------- [2019-04-24 14:47:48] Checking for Bowtie Bowtie version: 2.2.4.0 [2019-04-24 14:47:48] Checking for Bowtie index files (genome).. [2019-04-24 14:47:48] Checking for reference FASTA file [2019-04-24 14:47:48] Generating SAM header for Bowtie2Index/genome [2019-04-24 14:47:55] Reading known junctions from GTF file [2019-04-24 14:47:59] Preparing reads left reads: min. length=32, max. length=36, 21256592 kept reads (52962 discarded) right reads: min. length=32, max. length=36, 20996406 kept reads (313148 discarded) [2019-04-24 14:56:54] Building transcriptome data files /scratch/5555360.1.linga/tophat2/tmp/RefSeq_GeneBody [2019-04-24 14:57:14] Building Bowtie index from RefSeq_GeneBody.fa [2019-04-24 15:05:18] Mapping left_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-24 15:26:58] Mapping right_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-24 15:41:51] 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 15:41:51] Mapping left_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-24 15:46:26] Mapping right_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-24 15:51:29] Searching for junctions via segment mapping [2019-04-24 15:53:47] Retrieving sequences for splices [2019-04-24 15:56:08] Indexing splices [2019-04-24 15:56:46] 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/5555360.1.linga/tophat2/tmp/left_kept_reads.m2g_um_unmapped.bam [2019-04-24 15:56:54] Joining segment hits [2019-04-24 16:03:59] 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/5555360.1.linga/tophat2/tmp/right_kept_reads.m2g_um_unmapped.bam [2019-04-24 16:04:09] Joining segment hits [2019-04-24 16:10:20] Reporting output tracks ----------------------------------------------- [2019-04-24 16:45:03] A summary of the alignment counts can be found in /scratch/5555360.1.linga/tophat2/align_summary.txt [2019-04-24 16:45:03] Run complete: 01:57:14 elapsed