Error: Please supply a reference FASTA/GBK/EMBL file with --reference
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7 weeks ago
Neel ▴ 10

Hi, i am trying to run snippy on multiple genomes, however it gives following error like Please supply a reference FASTA/GBK/EMBL file with --reference even after providing the reference file. i don't understand why it is happen and here is code which i am using-

@ #

# specify the directory containing the genome files

genome_dir=/home/bvs/Neelam/fna

# specify the reference genome

ref=/home/bvs/Neelam/ref_snippy/PAO1.fna

# specify the directory for the output files

out_dir=/home/bvs/Neelam/snippy_outdir

# loop through all the genome files in the directory

for file in $(find$genome_dir -type f -name "*.fna"); do

# extract the sample name from the file name

name=$(echo$file | cut -d"/" -f 2 | cut -d"." -f1)

# run Snippy with the sample name as the prefix

snippy --prefix $name$file --cpus 30 --outdir $out_dir --reference$ref --ctgs $genome_dir done This is snippy 4.6.0 [16:47:05] Written by Torsten Seemann [16:47:05] Obtained from https://github.com/tseemann/snippy [16:47:05] Detected operating system: linux [16:47:05] Enabling bundled linux tools. [16:47:05] Found bwa - /usr/bin/bwa [16:47:05] Found bcftools - /home/bvs/Neelam/snippy/binaries/linux/bcftools [16:47:05] Found samtools - /usr/bin/samtools [16:47:05] Found java - /usr/bin/java [16:47:05] Found snpEff - /home/bvs/Neelam/snippy/binaries/noarch/snpEff [16:47:05] Found samclip - /home/bvs/Neelam/snippy/binaries/noarch/samclip [16:47:05] Found seqtk - /usr/bin/seqtk [16:47:05] Found parallel - /usr/bin/parallel [16:47:05] Found freebayes - /home/bvs/Neelam/snippy/binaries/linux/freebayes [16:47:05] Found freebayes-parallel - /home/bvs/Neelam/snippy/binaries/noarch/freebayes-parallel [16:47:05] Found fasta_generate_regions.py - /home/bvs/Neelam/snippy/binaries/noarch/fasta_generate_regions.py [16:47:05] Found vcfstreamsort - /home/bvs/Neelam/snippy/binaries/linux/vcfstreamsort [16:47:05] Found vcfuniq - /home/bvs/Neelam/snippy/binaries/linux/vcfuniq [16:47:05] Found vcffirstheader - /home/bvs/Neelam/snippy/binaries/noarch/vcffirstheader [16:47:05] Found gzip - /usr/bin/gzip [16:47:05] Found vt - /home/bvs/Neelam/snippy/binaries/linux/vt [16:47:05] Found snippy-vcf_to_tab - /home/bvs/Neelam/snippy/bin/snippy-vcf_to_tab [16:47:05] Found snippy-vcf_report - /home/bvs/Neelam/snippy/bin/snippy-vcf_report [16:47:05] Checking version: samtools --version is >= 1.7 - ok, have 1.9 [16:47:05] Checking version: bcftools --version is >= 1.7 - ok, have 1.10 [16:47:05] Checking version: freebayes --version is >= 1.1 - ok, have 1.3.1 [16:47:06] Checking version: snpEff -version is >= 4.3 - ok, have 4.3 [16:47:06] Checking version: bwa is >= 0.7.12 - ok, have 0.7.17 [16:47:06] **Please supply a reference FASTA/GBK/EMBL file with --reference** Thank you!  snippy Prokka • 384 views ADD COMMENT 0 Entering edit mode 7 weeks ago Type echo$ref

to see if your ref has been saved in an environment variable. If this is empty it is a problem.

Else reenter your line, then retry

ref=/home/bvs/Neelam/ref_snippy/PAO1.fna

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