[2019-04-24 14:54:52] Beginning TopHat run (v2.1.1) ----------------------------------------------- [2019-04-24 14:54:52] Checking for Bowtie Bowtie version: 2.2.4.0 [2019-04-24 14:54:53] Checking for Bowtie index files (genome).. [2019-04-24 14:54:53] Checking for reference FASTA file [2019-04-24 14:54:53] Generating SAM header for Bowtie2Index/genome [2019-04-24 14:54:57] Reading known junctions from GTF file [2019-04-24 14:55:02] Preparing reads left reads: min. length=32, max. length=36, 23963312 kept reads (114772 discarded) right reads: min. length=32, max. length=36, 23648689 kept reads (429395 discarded) [2019-04-24 15:05:04] Building transcriptome data files /scratch/5555369.1.linga/tophat2/tmp/RefSeq_GeneBody [2019-04-24 15:05:24] Building Bowtie index from RefSeq_GeneBody.fa [2019-04-24 15:14:14] Mapping left_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-24 15:27:17] Mapping right_kept_reads to transcriptome RefSeq_GeneBody with Bowtie2 [2019-04-24 15:41:17] 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 15:41:17] Mapping left_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-24 15:45:01] Mapping right_kept_reads.m2g_um to genome genome with Bowtie2 [2019-04-24 15:50:29] Searching for junctions via segment mapping [2019-04-24 15:52:31] Retrieving sequences for splices [2019-04-24 15:54:42] Indexing splices [2019-04-24 15:55:05] 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/5555369.1.linga/tophat2/tmp/left_kept_reads.m2g_um_unmapped.bam [2019-04-24 15:55:09] Joining segment hits [2019-04-24 16:00:37] 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/5555369.1.linga/tophat2/tmp/right_kept_reads.m2g_um_unmapped.bam [2019-04-24 16:00:41] Joining segment hits [2019-04-24 16:06:03] Reporting output tracks ----------------------------------------------- [2019-04-24 16:26:44] A summary of the alignment counts can be found in /scratch/5555369.1.linga/tophat2/align_summary.txt [2019-04-24 16:26:44] Run complete: 01:31:51 elapsed