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Repo for working on long-read triple amidite bulk RNA-seq

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TallyTriN

Overview

Long-read sequencing has become an increasingly popular tool for RNA sequencing that provides unprecedented insight into isoform, translocation and variant calling analysis.

Further documentation and tutorial on how to run a workflow can be found at read the docs here

TallyTriN is a collection of bulk and single-cell workflows that utilize Unique Molecular Identifiers (UMIs) synthesized using trimer blocks of nucleotides.

Workflows

Included within this repo are the follwing workflows:

  • pipeline_count - a bulk RNA-seq workflow that facilitates the analysis of UMIs synthesised using trimer nucleoside blocks added to the 5' and 3' of the RNA molecule.
  • pipeline_fusion - a bulk analysis pipeline for the analysis of fusion transcripts. It implements a strategy for removing chimeric artefacts and quantifies real genomic translocation at the RNA-seq level.
  • pipeline_singlecell - a workflow that facilitates the analysis of scCOLOR-seq nanopore single-cell sequencing data, but with trimers added at the 5' end of the RNA.
  • pipeline_10X - a workflow that facilitates the analysis of 10X sequencing data.

Installation

We reccomend installing miniconda, then creating a new environment and install mamba. This istallation typically takes 10 minutes.

conda install mamba -c conda-forge

Next install the required software using the conda yml file

# Clone the TallyTriN repo
git clone https://github.com/cribbslab/TallyTriN.git
# Use mamba to install environment
mamba env update --file conda/environment.yml

Activate the condda environment

conda activate tallytrin

Then, you will need to manually install TallyTriN and the fork of umi tools. The fork is added as a submodule to this repo to help you easily install.

# Install TallyTriN code
pip install -e .

Documentation

Further information how you can run the pipelines can be found at read the docs

Usage

Run the tallytrin --help command to see what workflows are available and tallytrin count -help to see how to use them.

For example, to generate a configuration file run

tallytrin count config

To set up the configuration file, please refer to our documentation.

To run the pipeline with all tasks, run

tallytrin count make full -v5 

Manuscript

The bioRxiv manuscript accompanying this code can be found here:

https://www.biorxiv.org/content/10.1101/2023.04.06.535911v1

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Repo for working on long-read triple amidite bulk RNA-seq

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