Hello All,
I am pretty new to RNA-Seq data analysis and trying to use Tophat for alignment of RNA-Seq reads data.
I am getting below error while running the Tophat Alignment job.
[2016-04-11 02:18:26] Beginning TopHat run (v2.1.0)
-----------------------------------------------
[2016-04-11 02:18:26] Checking for Bowtie
Bowtie version: 2.2.6.0
[2016-04-11 02:18:26] Checking for Bowtie index files (genome)..
[2016-04-11 02:18:26] Checking for reference FASTA file
Warning: Could not find FASTA file index/athal.fa
[2016-04-11 02:18:26] Reconstituting reference FASTA file from Bowtie index
Executing: /home/rajesh/Desktop/NGS_RNA_seq_Griffiths_lab/tools/bam-readcount/bin/bowtie2-2.2.6/bowtie2-inspect index/athal > ./tophat_out/tmp/athal.fa
[2016-04-11 02:18:26] Generating SAM header for index/athal
[2016-04-11 02:18:26] Preparing reads
WARNING: read pairing issues detected (check prep_reads.log) !
left reads: min. length=50, max. length=50, 57985 kept reads (0 discarded)
right reads: min. length=50, max. length=50, 57985 kept reads (0 discarded)
[2016-04-11 02:18:27] Mapping left_kept_reads to genome athal with Bowtie2
[2016-04-11 02:18:30] Mapping left_kept_reads_seg1 to genome athal with Bowtie2 (1/2)
[2016-04-11 02:18:30] Mapping left_kept_reads_seg2 to genome athal with Bowtie2 (2/2)
[2016-04-11 02:18:30] Mapping right_kept_reads to genome athal with Bowtie2
[2016-04-11 02:18:33] Mapping right_kept_reads_seg1 to genome athal with Bowtie2 (1/2)
[2016-04-11 02:18:33] Mapping right_kept_reads_seg2 to genome athal with Bowtie2 (2/2)
[2016-04-11 02:18:34] 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 =127
/usr/local/bin/segment_juncs: error while loading shared libraries: libboost_thread.so.1.48.0: cannot open shared object file: No such file or directory
- I can see that the segment_juncs file is present in the usr/local/bin directory.
Below are some of the solutions posted earlier which did not help in my case:
1) I have already tried to export the path in variable LD_LIBRARY_PATH (which doesn’t work).
2) Tried to re-install libboost_thread.so.1.48.0, however, getting the error "Couldn't find any package by regex 'libboost_thread.so.1.48.0"
I would really appreciate if anyone could suggest any possible workaround or provide the solution for this particular error.
Thanks in advance for your time and help.
rna-seq
tophat
alignment
libboost
segment_juncs