16S_workflow/16S_workflow.sh

#!/bin/bash
raw_reads_dir=$1
output_dir=$2

mkdir $output_dir
mkdir trimmed_reads
#TRIM READS WITH ALIENTRIMMER
for R1 in $(ls $raw_reads_dir/*R1.fastq);
    do
        R2=`echo $R1 | sed 's/R1/R2/g'`
        oR1=`echo $R1 | sed 's/\.fastq/\.at\.fq/g'`
        oR1=`echo $oR1 | sed 's/fastq\///g'`
        oR2=`echo $R2 | sed 's/\.fastq/\.at\.fq/g'`
        oR2=`echo $oR2 | sed 's/fastq\///g'`
        java -jar soft/AlienTrimmer.jar -if $R1 -ir $R2 -q 20 -c databases/contaminants.fasta -of trimmed_reads/$oR1 -or trimmed_reads/$oR2
    done


# MERGE READS AND OUTPUT FASTA
mkdir fasta
for R1 in $(ls trimmed_reads/*R1.at.fq);
    do
        R2=`echo $R1 | sed 's/R1/R2/g'`
        outname=`echo $R1 | sed 's/_R1\.at\.fq/\.fasta/g'`
        label=`echo $outname | sed 's/trimmed_reads\///g'`
        outname=`echo $outname | sed 's/trimmed_reads/fasta/g'`
        label=`echo $label | sed 's/\.fasta//g'`
        soft/vsearch --fastq_mergepairs $R1 --reverse $R2 --fastaout $outname --label_suffix ";sample=$label;"
    done

# MERGE ALL FASTAS TO 'amplicon.fasta'
cat fasta/*.fasta > fasta/amplicon.fasta

# REMOVE ' ' FROM EACH READ NAME
sed -i 's/ //g' fasta/amplicon.fasta

# CREATE OTU
# DEDUPLICATION
mkdir otu
soft/vsearch --derep_fulllength fasta/amplicon.fasta --output fasta/fulllength.fasta
soft/vsearch --derep_prefix fasta/amplicon.fasta --output fasta/prefix.fasta

# REMOVING SINGLETON
soft/vsearch --derep_prefix fasta/amplicon.fasta --output fasta/prefix_min10.fasta --minuniquesize 10

# REMOVING CHIMERAS
soft/vsearch --uchime_denovo fasta/prefix.fasta --nonchimeras fasta/prefix_nochim.fasta

# CLUSTERING
otu=">OTU_"
vsearch --cluster_fast fasta/prefix_nochim.fasta --id 0.97 --centroids otu/otu.fasta --relabel $otu

# ABONDANCE
vsearch --usearch_global fasta/amplicon.fasta --db otu/otu.fasta --id 0.97 --otutabout otu/abondance_table.txt

# ANNOTATION
soft/vsearch --usearch_global otu/otu.fasta --db databases/mock_16S_18S.fasta --id 0.97 --top_hits_only --userfields query+target --userout otu/annotation.txt