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Error with hera not finding the index

Dear all,

I have created a Hera index using:

./output/software/hera/build/hera_build --fasta output/genome/celegans/ref/seq/celegans.fa --gtf output/genome/celegans/ref/annotation/celegans.gtf --outdir output/genome/celegans/ref/index/hera/

This creates 3 files: index, reference.inf, transcripts.fasta

I then use:

./output/software/hera/build/hera quant -i output/genome/celegans/ref/index/hera/  output/reads/illumina/trimmed/pooled/altadena1.fastq

but this creates the following error:

Can not open index file or file is not exist!!!

Can anybody help me, please?

Thanks.

Best,

C.

hera index error

Have you checked hera help? It looks like you may need to include the basename for the index. Instead of -i output/genome/celegans/ref/index/hera/ e.g. -i output/genome/celegans/ref/index/hera/genome.

Hi,

I tried your change but it did not work. Why two forward slashes? And why 'genome' in particular? What is the basename?

From what is written here: https://github.com/bioturing/hera I cannot find anything about the basename.

Thanks.

Best,

C.

Double-slashes were a typo. Fixed.

Perhaps another recommendation for the developers would be to change that error message from:

Can not open index file or file is not exist!!!

To:

Can not open index file or file does not exist!!!

Thanks WouterDeCoster. We have fixed the typos but still can't reproduce the error!

Hi Cristian, Thank you for the bug report and sorry for the error! Can you tell me what is the output of "ls output/genome/celegans/ref/index/hera/"

Hi,

Here it is.

ls output/genome/celegans/ref/index/hera/
index             reference.inf     transcripts.fasta

Best, C.

Hi cristian, I have created a similar working directory as your command and tried to map a fastq read file on c.elegans genome (from ensembl). It turned out to work normally on my system.

./output/software/hera/build/hera_build --fasta output/genome/celegans/ref/seq/Caenorhabditis_elegans.WBcel235.dna_sm.toplevel.fa --gtf output/genome/celegans/ref/annotation/Caenorhabditis_elegans.WBcel235.89.gtf --outdir output/genome/celegans/ref/index/hera/
Scan reference sequence ...
Scan gtf file...
Extract transcript sequence
Hera is a program developed by BioTuring for RNA-Seq analysis.
Please contact info@bioturing.com if you need further support
Index transcripts in: output/genome/celegans/ref/index/hera/transcripts.fasta
Number of sequence  : 58941
____

ls -lha output/genome/celegans/ref/index/hera/
total 1,1G
drwxrwxr-x 2 thaottp thaottp 4,0K Th07 30 22:42 .
drwxrwxr-x 3 thaottp thaottp 4,0K Th07 30 22:38 ..
-rw-rw-r-- 1 thaottp thaottp 973M Th07 30 22:42 index
-rw-rw-r-- 1 thaottp thaottp 3,9M Th07 30 22:41 reference.inf
-rw-rw-r-- 1 thaottp thaottp  54M Th07 30 22:41 transcripts.fasta
___

./output/software/hera/build/hera quant -i output/genome/celegans/ref/index/hera/ sim/sim1_1.fq 
Hera is a program developed by BioTuring for RNA-Seq analysis.
Please contact info@bioturing.com if you need further support
Process single-end reads in: sim/sim1_1.fq
Number of aligned readsd: 1692/30000000000      
Mean read lenght    : 75.000000
Finish EM with 50 rounds

Can you give me more details about your system (uname -a) and results from index step (ls -lha output/genome/celegans/ref/index/hera/)?

Hi phuongthao.12csh,

Since your reaction is a query for more information and not (yet) an answer to OPs question I have moved your reaction to a comment rather than an answer.

Cheers,
Wouter

Hi, Thanks.

uname -a
Darwin cr517s-MacBook-Pro.local 16.6.0 Darwin Kernel Version 16.6.0: Fri Apr 14 16:21:16 PDT 2017; root:xnu-3789.60.24~6/RELEASE_X86_64 x86_64

Best, C.

And,

ls -lha output/genome/celegans/ref/index/hera/ 
total 2109200
drwxr-xr-x  6 cr517  staff   204B 30 Jul 16:22 .
drwxr-xr-x  7 cr517  staff   238B 30 Jul 14:33 ..
-rw-r--r--@ 1 cr517  staff   6.0K 30 Jul 16:22 .DS_Store
-rw-r--r--  1 cr517  staff   973M 30 Jul 16:22 index
-rw-r--r--  1 cr517  staff   3.8M 30 Jul 16:22 reference.inf
-rw-r--r--  1 cr517  staff    53M 30 Jul 16:22 transcripts.fasta

1 answer

Hi cristian,

It seems that the error is due to the difference between Mac and Linux. We have fixed it and committed to Github. Please pull or clone ( https://github.com/bioturing/hera ), and check if it can work now.

Bioturing Algorithm Team.

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