New algorithm for full viral haplotype reconstruction and abundance estimation. It is an alternative approach for the first developed approach viaDBG which was based entirely on de Bruijn graphs. ViQUF is based on flow networks which allows us to do a proper and successful estimation of the strains frequencies.
The overall workflow is as follows:
- Building assembly graph (BCALM)
- Polishing the assembly graph by:
- Classifying edges as weak and strong, and removing the weak ones called as filigree edges.
- Removing isolated nodes.
- Removing short tips.
- Paired-end association.
- Paired-end polishing removing as many wrong associations as possible.
- Core algorithm:
- For every pair of adjacent nodes A -> B, we built a DAG from their paired-end information.
- DAG is translated into a flow network and a min-cost flow is solved.
- The flow is translated into paths via a "greedy" path heuristic.
- Final strains are build following two rules:
- Standard contig traversion (deprecated)
- Min-cost flow over the Approximate Paired de Bruijn Graph built from the core algorithm.
- Python 3.* - we encourage you to build a conda environment and still all dependencies via conda: conda create -n ViQUF-env python=3.6
- Biopython, altair, gurobi
- matplotlib, scipy, numpy
- C++17
- SDSL
- BCALM
- quast 4.3 or quast 5.0.1 to evaluate the results
- gatb-library:
- cd lib/ && rm -r gatb-core
- git clone https://github.com/GATB/gatb-core.git
- follow the instructions in: https://github.com/GATB/gatb-core
- lemon 1.3.1:
- cd ..
- wget http://lemon.cs.elte.hu/pub/sources/lemon-1.3.1.tar.gz
- tar xvf lemon-1.3.1.tar.gz
- follow instructions in https://lemon.cs.elte.hu/trac/lemon/wiki/InstallLinux (IMPORTANT: install lemon in /usr/local otherwise the Makefile will not get the lemon package)
- Compile: make clean && make
- Amplicons - new approach to deal with amplicons (On progress).
- Third Generation Sequencing - new approaches to deal with this type of data. Right now, we are able to infer a valid flow from the data (all test over simlord high depth simulated data).
- TODO: Apply Flow decomposition with subpath constraints.
Exists a Dockerfile which automatizes the installation procedure. To use it just run:
- sudo docker rm [your_decision_docker_name]
- sudo docker build -t [your_decision_docker_name] . --no-cache
- sudo docker run -d --name [your_decision_docker_name] [your_decision_docker_name]
The file execution-script contains an example about how to execute the code.
- python scripts/testBcalm.py $1 $2 10 ngs $3 $4 --no-meta
- $1 - folder with NGS reads
- $2 - kmer size
- $3 - --correct/--no-correct to perform correction or not respectively
- $4 - --joined, has pear been executed? If so --joined otherwise --no-join
- ./bin/output.out tmp $2 tmp/unitigs.graph tmp/unitigs.unitigs.fa tmp/unitigs-viadbg.fa tmp/Ownlatest/append.fasta $5 $6 --virus
- $5 - complete set of reads (it is not mandatory but recommended)
- $6 - --debug or not.
- python scripts/post-process.py
- Linear programming algorithm to adjust contigs frequencies, there is not mandatory but suggested.