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MinION_longamp

tools for long amplicon data processing for bacterial communities sequenced on a MinION

This workflow is used for obtaining consensus sequences and a count table, starting from demultiplexed fastq files. This can be done in a single step using workflow_all_in_one.sh or in multiple seperate steps (see below). ##Installation: This requires no additional software apart from what is already required to run NGSpeciesID. you can find the instructions to install NGSpeciesID here. Copy the required scipts provided in this branch to the folder containing the fasq files that you want to analyse.

workflow_all_in_one.sh can be executed from the folder that contains the fastq files. It accepts one optional argument, the abundance ratio cutoff for NGSpeciesID. if this is not given, it is 0.002

bash workflow_all_in_one.sh

the same workflow can be run in different steps to increase flexiblity

1 add sample code to header
bash add_sample_code.sh

ls *fastq | sed -e 's/\.fastq$//' >file_names.txt

2 merge fastqs if relevant cat *fastq > grouped.fastq

3 run NGSpeciesID ./NGSpeciesID --ont --fastq file.fastq --consensus --abundance_ratio 0.002 --racon --racon_iter 2 --outfolder folder_out

4 count number of reads per sample per consensus seq

bash readcount.sh ../file_names.txt

6 merge concensus sequences cat *fasta >all_consensus.fasta

taxonomic assignment ...