Hello,

I don't know why I still have this problem..

I have run tophat2 with different dataset, sometimes it goes well but sometime I have this error.

I run only one job at a time on a virtual machine with 8G memory without using galaxy plateform. I tried --no-coverage-search option but it changes nothing.

Thanks.

Delong

De : Delong, Zhou
Envoyé : 27 août 2013 9:36
À : galaxy-user@bx.psu.edu
Objet : Tophat Error: segment-based junction search failed with err

Hello,

I have run several analysis with Tophat 2 on my local instance of galaxy and I get this error for all of them..

segment-based junction search failed with err = 1 or -9

Here is an example of full error report:

Error in tophat:

[2013-08-23 11:56:58] Beginning TopHat run (v2.0.6)
-----------------------------------------------
[2013-08-23 11:56:58] Checking for Bowtie
		  Bowtie version:	 2.0.2.0
[2013-08-23 11:56:58] Checking for Samtools
		Samtools version:	 0.1.18.0
[2013-08-23 11:56:58] Checking for Bowtie index files
[2013-08-23 11:56:58] Checking for reference FASTA file
[2013-08-23 11:56:58] Generating SAM header for /usr/local/data/bowtie2/hg19/hg19
	format:		 fastq
	quality scale:	 phred33 (default)
[2013-08-23 11:58:04] Preparing reads
	 left reads: min. length=50, max. length=50, 145339247 kept reads (34946 discarded)
	right reads: min. length=50, max. length=50, 145340153 kept reads (34040 discarded)
[2013-08-23 14:16:21] Mapping left_kept_reads to genome hg19 with Bowtie2 
[2013-08-24 01:04:37] Mapping left_kept_reads_seg1 to genome hg19 with Bowtie2 (1/2)
[2013-08-24 03:38:22] Mapping left_kept_reads_seg2 to genome hg19 with Bowtie2 (2/2)
[2013-08-24 05:29:58] Mapping right_kept_reads to genome hg19 with Bowtie2 
[2013-08-24 19:50:22] Mapping right_kept_reads_seg1 to genome hg19 with Bowtie2 (1/2)
[2013-08-24 22:36:38] Mapping right_kept_reads_seg2 to genome hg19 with Bowtie2 (2/2)
[2013-08-25 01:40:37] Searching for junctions via segment mapping
	Coverage-search algorithm is turned on, making this step very slow
	Please try running TopHat again with the option (--no-coverage-search) if this step takes too much time or memory.
	[FAILED]
Error: segment-based junction search failed with err =-9
Collecting potential splice sites in islands


cp: cannot stat `/home/galaxy/galaxy-dist/database/job_working_directory/000/515/tophat_out/deletions.bed': No such file or directory
cp: cannot stat `/home/galaxy/galaxy-dist/database/job_working_directory/000/515/tophat_out/insertions.bed': No such file or directory

I did some research on the internet and it seems to be a memory problem to me, is there any solution other than rerun these jobs on a more powerful machine?

And why has Bowtie/Tophat discard different numbers of reads? What will be the impact? Does it means that if I don't have exact matches between the paired end input, it is still be possible to run the job?

Thanks,

Delong