Skip to content
New issue

Have a question about this project? Sign up for a free GitHub account to open an issue and contact its maintainers and the community.

By clicking “Sign up for GitHub”, you agree to our terms of service and privacy statement. We’ll occasionally send you account related emails.

Already on GitHub? Sign in to your account

fail to open "accepted_hits.bam" for reading; Error reading _plen[] array: 890800743, 4294967292 #10

Open
lokdeep1993 opened this issue Mar 18, 2019 · 0 comments

Comments

@lokdeep1993
Copy link

lokdeep1993 commented Mar 18, 2019

Hello,
I am trying to run UROBORUS on reads simulated from human cdna of length 150bp.
I used this link to download UROBORUS:
wget https://github.com/WGLab/UROBORUS/archive/v2.0.0.tar.gz
I used TopHat2 and Bowtie2 to generate unmapped.bam and used samtools (Version: 0.1.19-44428cd) to generate unmapped.sam.
Then, I tried to run UROBORUS on unmapped.sam using command:
uro_path=/home/nagarjun/Downloads/UROBORUS-2.0.0/bin index_path=/data2/circrna/bow1index perl "$uro_path"/UROBORUS.pl -index "$index_path"/humbow1 -gtf "$index_path"/human.gtf -fasta "$index_path" unmapped.sam accepted_hits.bam

Then, I started to get series of errors; and the final circRNA list is empty.

[nagarjun@localhost tophat_out]$ perl "$uro_path"/UROBORUS.pl -index "$index_path"/humbow1 -gtf "$index_path"/human.gtf -fasta "$index_path" unmapped.sam accepted_hits.bam
[Mon Mar 18 14:26:05 2019] Preparing the reads..........finished!
[Mon Mar 18 14:26:08 2019] Now tophat..........
[2019-03-18 14:26:08] Beginning TopHat run (v2.1.1)

[2019-03-18 14:26:08] Checking for Bowtie
Bowtie version: 1.2.2.0
[2019-03-18 14:26:08] Checking for Bowtie index files (genome)..
[2019-03-18 14:26:08] Checking for reference FASTA file
[2019-03-18 14:26:08] Generating SAM header for /data2/circrna/bow1index/humbow1
[2019-03-18 14:26:09] Preparing reads
left reads: min. length=20, max. length=20, 489200 kept reads (56 discarded)
right reads: min. length=20, max. length=20, 489206 kept reads (50 discarded)
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
[2019-03-18 14:26:14] Mapping left_kept_reads to genome humbow1 with Bowtie
[FAILED]
Error running bowtie:
Error reading _plen[] array: 890800743, 4294967292
Command: /usr/bin/bowtie-align-s --wrapper basic-0 -v 2 -k 20 -m 20 -S -p 6 --sam-nohead --max /dev/null /data2/circrna/bow1index/humbow1 -

finished!
open: No such file or directory
[main_samview] fail to open "accepted_hits.bam" for reading.
[Mon Mar 18 14:26:14 2019] Filtering the mapped seeds...........finished!
[Mon Mar 18 14:26:16 2019] Locating the seeds in the genome...........finished!
[Mon Mar 18 14:26:16 2019] Locating the seed lab ...........finished!
[Mon Mar 18 14:26:16 2019] Extending the paired seeds...........[Mon Mar 18 14:26:16 2019]
finished!
[Mon Mar 18 14:26:16 2019] Extending the single seed...........[Mon Mar 18 14:26:16 2019]
finished!
[Mon Mar 18 14:26:16 2019] Now bowtie..........Warning: -M was specified w/o --best; automatically enabling --best
Error reading _plen[] array: 890800743, 4294967292
Command: /usr/bin/bowtie-align-s --wrapper basic-0 -p 6 -n 0 -M 20 -I 100 -X 100000 --ff -1 IMR_1.fastq -2 IMR_2.fastq -S --sam-nohead /data2/circrna/bow1index/humbow1
finished!
[Mon Mar 18 14:26:16 2019]
[Mon Mar 18 14:26:16 2019] Now filtering mapped IMR..........finished!
[Mon Mar 18 14:26:16 2019]
[Mon Mar 18 14:26:16 2019] Now sorting mapped IMR sam file..........Use of uninitialized value $line in split at /home/nagarjun/Downloads/UROBORUS-2.0.0/bin/UROBORUS.pl line 1561.
Use of uninitialized value $line in split at /home/nagarjun/Downloads/UROBORUS-2.0.0/bin/UROBORUS.pl line 1564.
Use of uninitialized value $out_line in print at /home/nagarjun/Downloads/UROBORUS-2.0.0/bin/UROBORUS.pl line 1580.
finished!
[Mon Mar 18 14:26:16 2019] Now locating mapped IMR..........finished!
[Mon Mar 18 14:26:16 2019]
[Mon Mar 18 14:26:16 2019] Now mating the reads...........finished!
[Mon Mar 18 14:26:16 2019]
[Mon Mar 18 14:26:16 2019] Now filtering the accepted reads...........open: No such file or directory
[bam_sort_core] fail to open file accepted_hits.sorted.bam
open: No such file or directory
[main_samview] fail to open "accepted_hits.sorted.bam" for reading.
Use of uninitialized value $1 in print at /home/nagarjun/Downloads/UROBORUS-2.0.0/bin/UROBORUS.pl line 1778.
Use of uninitialized value $1 in print at /home/nagarjun/Downloads/UROBORUS-2.0.0/bin/UROBORUS.pl line 1778.
Use of uninitialized value $1 in print at /home/nagarjun/Downloads/UROBORUS-2.0.0/bin/UROBORUS.pl line 1778.
Use of uninitialized value $1 in print at /home/nagarjun/Downloads/UROBORUS-2.0.0/bin/UROBORUS.pl line 1778.
Use of uninitialized value $1 in print at /home/nagarjun/Downloads/UROBORUS-2.0.0/bin/UROBORUS.pl line 1778.
Use of uninitialized value $1 in print at /home/nagarjun/Downloads/UROBORUS-2.0.0/bin/UROBORUS.pl line 1778.
Use of uninitialized value $1 in print at /home/nagarjun/Downloads/UROBORUS-2.0.0/bin/UROBORUS.pl line 1778.
Use of uninitialized value $1 in print at /home/nagarjun/Downloads/UROBORUS-2.0.0/bin/UROBORUS.pl line 1778.
Use of uninitialized value $1 in print at /home/nagarjun/Downloads/UROBORUS-2.0.0/bin/UROBORUS.pl line 1778.
[Mon Mar 18 14:27:12 2019]
[Mon Mar 18 14:27:12 2019] Merging the results...........finished!
[Mon Mar 18 14:27:12 2019] Making the statistics...........finished!
[Mon Mar 18 14:27:12 2019] Output the circular RNA list!
[Mon Mar 18 14:27:12 2019] Completed successfully! The time elapsed: about 0.01 hours.

-rw-rw-r-- 1 nagarjun nagarjun 188 Mar 16 15:04 prep_reads.info
-rw-rw-r-- 1 nagarjun nagarjun 563 Mar 16 23:30 align_summary.txt
-rw-rw-r-- 1 nagarjun nagarjun 421592 Mar 16 23:30 insertions.bed
-rw-rw-r-- 1 nagarjun nagarjun 874653 Mar 16 23:30 deletions.bed
-rw-rw-r-- 1 nagarjun nagarjun 26194280 Mar 16 23:30 junctions.bed
-rw-rw-r-- 1 nagarjun nagarjun 7520512819 Mar 17 00:34 accepted_hits.bam
drwxrwxr-x 2 nagarjun nagarjun 4096 Mar 17 00:34 logs
-rw-rw-r-- 1 nagarjun nagarjun 49666683 Mar 17 00:35 unmapped.bam
-rw-rw-r-- 1 nagarjun nagarjun 167132556 Mar 18 11:26 unmapped.sam
drwxrwxr-x 4 nagarjun nagarjun 97 Mar 18 14:26 seed_mapped_out
-rw-rw-r-- 1 nagarjun nagarjun 0 Mar 18 14:26 accepted_hits.sorted.sam
-rw-rw-r-- 1 nagarjun nagarjun 0 Mar 18 14:26 PI_list.txt
drwxrwxr-x 3 nagarjun nagarjun 29 Mar 18 14:26 tmp_3_18_14_26
-rw-rw-r-- 1 nagarjun nagarjun 0 Mar 18 14:27 circRNA_list.txt`

Please help.
I also want to mention that initially it was giving error of not finding genome index file with humbow1.*.ebwt
I checked and found that the indexed files have extension *.ebwtl
I renamed the files to *.ebwt and the error was gone. IS this the right approach?
Thank You in advance

Sign up for free to join this conversation on GitHub. Already have an account? Sign in to comment
Labels
None yet
Projects
None yet
Development

No branches or pull requests

1 participant