Question: TopHat Fata Error: Fusion search fails with Bowtie2, how to try Bowtie1 option? Also truncates file during genome alignment
0
gravatar for marissasp
2.8 years ago by
marissasp0
marissasp0 wrote:

I am trying to run TopHat using a GTF annotation to build a transcriptome index, and also search for fusions.  The data becomes truncated during the second alignment step, to the reference genome.  I also receive a warning to try Bowtie 1 not Bowtie 2 when looking for fusions, but am not sure how to do this within TopHat.  The error message is: 

Fatal error: Tool execution failed
	Warning: --fusion-search with Bowtie2 may not work well as it may require much memory space and produce many spurious fusions.  Please try --bowtie1 option if this doesn't work.

[2016-02-15 12:47:05] Beginning TopHat run (v2.0.14)
-----------------------------------------------
[2016-02-15 12:47:05] Checking for Bowtie
		  Bowtie version:	 2.2.5.0
[2016-02-15 12:47:05] Checking for Bowtie index files (genome)..
[2016-02-15 12:47:05] Checking for reference FASTA file
[2016-02-15 12:47:05] Generating SAM header for /galaxy/data/hg19/hg19full/bowtie2_index/hg19full
[2016-02-15 12:47:07] Reading known junctions from GTF file
[2016-02-15 12:47:53] Preparing reads
	 left reads: min. length=100, max. length=100, 93564119 kept reads (18701 discarded)
	right reads: min. length=100, max. length=100, 93392573 kept reads (190247 discarded)
[2016-02-15 14:01:14] Building transcriptome data files ./tophat_out/tmp/dataset_14372302
[2016-02-15 14:03:11] Building Bowtie index from dataset_14372302.fa
[2016-02-15 16:08:31] Mapping left_kept_reads to transcriptome dataset_14372302 with Bowtie2 
[2016-02-16 01:15:00] Mapping right_kept_reads to transcriptome dataset_14372302 with Bowtie2 
[2016-02-16 10:14:43] Resuming TopHat pipeline with unmapped reads
[2016-02-16 10:14:43] Mapping left_kept_reads.m2g_um to genome hg19full with Bowtie2 
[main_samview] truncated file.
[main_samview] truncated file.
[main_samview] truncated file.
[2016-02-16 12:24:42] Mapping left_kept_reads.m2g_um_seg1 to genome hg19full with Bowtie2 (1/4)
[2016-02-16 12:34:48] Mapping left_kept_reads.m2g_um_seg2 to genome hg19full with Bowtie2 (2/4)
[2016-02-16 12:48:29] Mapping left_kept_reads.m2g_um_seg3 to genome hg19full with Bowtie2 (3/4)
[2016-02-16 13:02:08] Mapping left_kept_reads.m2g_um_seg4 to genome hg19full with Bowtie2 (4/4)
[2016-02-16 13:16:45] Mapping right_kept_reads.m2g_um to genome hg19full with Bowtie2 
[main_samview] truncated file.
[bam_header_read] EOF marker is absent. The input is probably truncated.
[bam_header_read] invalid BAM binary header (this is not a BAM file).
[main_samview] fail to read the header from "./tophat_out/tmp/right_kept_reads.m2g_um_unmapped.bam".
[2016-02-16 16:25:57] Searching for junctions via segment mapping
	[FAILED]
Error: segment-based junction search failed with err =1
*** glibc detected *** /galaxy-repl/main/deps/tophat/2.0.14/iuc/package_tophat_2_0_14/536f7bb5616d/bin/segment_juncs: double free or corruption (fasttop): 0x0000000002209010 ***  Any help is greatly appreciated, thank you very much!!!!

 

 

 

 

tophat • 1.8k views
ADD COMMENTlink modified 2.8 years ago • written 2.8 years ago by marissasp0
0
gravatar for Jennifer Hillman Jackson
2.8 years ago by
United States
Jennifer Hillman Jackson25k wrote:

Hello,

If this is occurring at http://usegalaxy.org, please try to execute the job again. There have been service issues that impact jobs.

Thanks, Jen, Galaxy team

 

ADD COMMENTlink written 2.8 years ago by Jennifer Hillman Jackson25k
0
gravatar for marissasp
2.8 years ago by
marissasp0
marissasp0 wrote:

OK thank you!  I'm running it again now.  

ADD COMMENTlink written 2.8 years ago by marissasp0
0
gravatar for marissasp
2.8 years ago by
marissasp0
marissasp0 wrote:

Hi Jen,

Thank you so much for your help!  It has failed another three times.  On the first attempt, TopHat failed when re-run, with an error message saying it had run longer than the maximum allowed run time.  I then tried running it using a smaller GTF (iGenomes in the Shared Data folder on Main, rather than the larger GENCODE GTF I had already tried using).  That run also failed, with an error message that the run used more memory than was allocated.  I then tried to run it again, using the smaller iGenomes GTF but not asking it to search for fusions.  This also failed, due to running longer than the maximum allowed run time.  What would you recommend trying?  Thank you!!  - Marissa Spino

 

 

ADD COMMENTlink written 2.8 years ago by marissasp0

Hello - For failures concerning memory, moving to a local or cloud Galaxy with more dedicated compute is the solution. For failures concerning walltime (job ran longer than the maximum allowed), the option to use the longer running cluster Stampede can be used (See the tool form option "Job Resource Parameters"). If the job still fails for walltime at Stampede, then a local or cloud Galaxy is the solution.

Section 2.8 of the Galaxy support wiki explains alternatives for working with data/jobs that exceed the compute resources at http://usegalaxy.org:
https://wiki.galaxyproject.org/Support

Thanks, Jen

ADD REPLYlink written 2.8 years ago by Jennifer Hillman Jackson25k
0
gravatar for marissasp
2.8 years ago by
marissasp0
marissasp0 wrote:

Thanks!  Re-running with TACC Stampede now.

ADD COMMENTlink written 2.8 years ago by marissasp0
0
gravatar for marissasp
2.8 years ago by
marissasp0
marissasp0 wrote:

I've tried running it on Stampede twice, it's failed both times with the error message "Remote job server indicated a problem running or monitoring this job".  Is TACC the correct version of Stampede for this?  Thank you!!

 

ADD COMMENTlink written 2.8 years ago by marissasp0
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