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MinION nanopore sequencing and assembly of a complete human papillomavirus genome: bioinformatics pipeline

A pipeline developed to analyse MinION sequencing data for the reconstruction of viral genomes

Description

This pipeline was developed to analyse MinION sequencig raw data, to generate a consensus sequence for a viral genome. This script perform a polishing of the raw data and generate a de novo assemblig using Canu tool. After, it takes the canu contigs and performs a step of polishing using Medaka tool.

Prerequisites

This pipe was written in python3 Guppy is required for the basecalling. Nanofilt and filtlong are required for the filtering of the reads. Canu is required for the assembling step. Medaka is required for the polishing step.

Installation

To run these scripts, each of the tools used needs to be installed before to launch the process. Tools to install: Guppy, Nanofilt, Filtlong, Canu, Medaka.
Get Guppy from the MinION comunity and install it

conda install -c bioconda nanofilt

conda install -c bioconda filtlong

conda install -c bioconda canu

Parameters

  • Mandatory

Name Example value Description
--fast5_path path/to/file path to fast5 files
--flowcell flowcell code the code of flowcell used (ex. FLO-MIN106)
--kit kit_used the MinION kit used (ex. SQK-LSK109)
--threads nr. of threads (int) threads to be used (ex. 8)
--num num_callers (int) number of callers to use (ex. 8)
--barcode barcode kit code the code of the barcodes kit used (ex. EXP-PBC001 )
--medaka_m medaka model define a model based on the basecaller (ex. r941_min_high_g303) - see medaka tool for more details -
  • Flags

Flags are special parameters without value.

Name Description
-h Display help

Usage

Part5:

python3 MinION_pipe.py fast5_path flowcell kit threads num barcode medaka_m 

Contributions

Name Email Description
Rosario Nicola Brancaccio rosariobrancaccio@yahoo.it Developer to contact for support
Massimo Tommasino tommasinom@iarc.fr
Tarik Gheit gheitt@iartc.fr

Authors

Rosario Nicola Brancaccio - (https://github.com/BioRB/)

License

GNU General Public License, version 3

References

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