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Differential analysis of RNA modifications using Oxford Nanopore Technologies (ONT) direct RNA sequencing data

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Differential RNA modification calling at the isoform resolution using Nanopore direct RNA sequencing

Docs: https://shimlab.github.io/mako

Nextflow Docker Singularity

Introduction

Mako is a bioinformatics pipeline designed for differential RNA modification calling at the isoform resolution using Nanopore direct RNA sequencing. It takes a samplesheet and output from Dorado and/or m6Anet, and applies various statistical methods to identify differentially modified sites between experimental conditions.

Mako will also produce interactive visualisations for quality control and assessment of sites through the makoview tool.

Steps

  1. Sample and read QC
  2. Site-level aggregation, filtering, and selection
  3. Choice of differential analysis methods:
    1. Linear mixed-effects models
    2. Modkit
  4. Visualization of results via makoview

mako pipeline diagram

Documentation

Please refer to the documentation for detailed instructions on how to use the pipeline, including input requirements, parameter settings, and output interpretation.

Installation

Read the quick start. But if you're in a rush:

# Clone the repository
git clone https://github.com/shimlab/mako && cd mako

# Test the pipeline
nextflow run main.nf --help
nextflow run main.nf -profile testing

Citations

Mako does not have a preprint yet. Please check back later for updates.

Contributors

This package is developed and maintained by Yulin Wu, Oliver Cheng, Sophie Wharrie and Heejung Shim in the Shim Lab at the University of Melbourne.

Please leave an issue if you find a bug or have a feature request. Thank you!

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