Current Version: 0.3.1
Release date: 12 October 2016
Important
GraphMap's command line has changed significantly between version 0.3.x and 0.2x - although many options remain similar, the usage is incompatible with the older releases due to explicit tool specification.
The first parameter is now mandatory, and specifies whether the mapping/alignment (./graphmap align) or overlapping (./graphmap owler) should be used.
For a detailed change log from the previous release, take a look at doc/changelog.md.
For more information on overlapping, take a look at overlap.md.
For detailed installation instructions, take a look at INSTALL.md file.
Description of custom parameters in GraphMap's SAM output can be found at doc/sam_output.md.
- Mapping position agnostic to alignment parameters.
- Consistently very high sensitivity and precision across different error profiles, rates and sequencing technologies even with default parameters.
- Circular genome handling to resolve coverage drops near ends of the genome.
- E-value.
- Meaningful mapping quality.
- Various alignment strategies (semiglobal bit-vector and Gotoh, anchored).
- Overlapping of reads for de novo assembly.
- ...and much more.
GraphMap is also used as an overlapper in a new de novo genome assembly project called Ra (https://github.com/mariokostelac/ra-integrate).
Ra attempts to create de novo assemblies from raw nanopore and PacBio reads without requiring error correction, for which a highly sensitive overlapper is required.
git clone https://github.com/isovic/graphmap.git
cd graphmap
make modules
make
# To align:
./bin/Linux-x64/graphmap align -r reference.fa -d reads.fasta -o output.sam
# To overlap:
./bin/Linux-x64/graphmap owler -r reads.fasta -d reads.fasta -o output.mhap
GraphMap is a novel mapper targeted at aligning long, error-prone third-generation sequencing data.
It is designed to handle Oxford Nanopore MinION 1d and 2d reads with very high sensitivity and accuracy, and also presents a significant improvement over the state-of-the-art for PacBio read mappers.
GraphMap was also designed for ease-of-use: the default parameters can handle a wide range of read lengths and error profiles, including: Illumina, PacBio and Oxford Nanopore.
This is an especially important feature for technologies where the error rates and error profiles can vary widely across, or even within, sequencing runs.
The GraphMap algorithm is structured to achieve high-sensitivity and speed using a five-stage read-funneling approach. In stage I, GraphMap uses a novel adaptation of gapped spaced seeds to efficiently reduce the search space and get seed hits as a form of coarse alignment. These are then refined in stage II using graph-based vertex-centric processing of seeds to efficiently construct alignment anchors. GraphMap then chains anchors using a kmer version of longest common subsequence (LCSk) construction (stage III), refines alignments by chaining anchors in the anchored mode or with a form of L1 linear regression in the semiglobal alignment mode (stage IV) and finally evaluates the remaining candidates to select the best location to reconstruct a final alignment (stage V). GraphMap computes a BLAST-like E-value as well as a mapping quality for its alignments.
Evaluation on MinION sequencing datasets against short and long-read mappers indicates that GraphMap increases mapping sensitivity by at least 15-80%. GraphMap alignments are the first to demonstrate consensus calling with <1 error in 100,000 bases, variant calling on the human genome with 76% improvement in sensitivity over the next best mapper (BWA-MEM), precise detection of structural variants from 100bp to 4kbp in length and species and strain-specific identification of pathogens using MinION reads.
Further details about the algorithm, comparison with other mappers and usage applications can be found in the preprint of our paper:
Fast and sensitive mapping of error-prone nanopore sequencing reads with GraphMap
Nanopore sequencing data of E. Coli UTI89 generated in-house and used in the paper now available on ENA:
PRJEB9557
To build GraphMap from source type:
make modules # This pulls the latest version of all required submodules
make
You will need a recent GCC/G++ version (>=4.7).
More installation instructions can be found in the INSTALL.md file.
# **Align** all reads from a given FASTA/FASTQ file using anchored alignment approach:
./graphmap align -r escherichia_coli.fa -d reads.fastq -o alignments.sam
# **Overlap** all reads from a given FASTA/FASTQ file and report overlaps in MHAP format (fast):
./graphmap owler -r reads.fa -d reads.fa -o overlaps.mhap
# Align all reads from a given FASTA/FASTQ file with default number of threads using semiglobal bit-vector alignment:
./graphmap align -a sg -r escherichia_coli.fa -d reads.fastq -o alignments.sam
# Overlap all reads from a given FASTA/FASTQ in a full GraphMap mode with generating alignments (slow):
./graphmap align -x overlap -r reads.fa -d reads.fa -o overlaps.sam
# Align reads using the Gotoh for semiglobal alignment:
./graphmap align -a sggotoh -r escherichia_coli.fa -d reads.fastq -o alignments.sam
# Align reads using Gotoh alignment with anchored approach:
./graphmap align -a anchorgotoh -r escherichia_coli.fa -d reads.fastq -o alignments.sam
# Process reads from a circular genome:
./graphmap align -C -r escherichia_coli.fa -d reads.fastq -o alignments.sam
# Threshold the E-value of alignments to 1e-100. Alignments with E-value > 1e-100 will be called unmapped:
./graphmap align --evalue 1e-100 -r escherichia_coli.fa -d reads.fastq -o alignments.sam
# Output all secondary alignments instead of only one best:
./graphmap align --secondary -r escherichia_coli.fa -d reads.fastq -o alignments.sam
# Control the similarity for secondary alignments. All alignments to within F*num_covered_bases from the best will be output.
./graphmap align --secondary -F 0.05 -r escherichia_coli.fa -d reads.fastq -o alignments.sam
# Limit the number of threads to 8, and load reads in batches of 50MB:
./graphmap align -t 8 -B 50 -r escherichia_coli.fa -d reads.fastq -o alignments.sam
# Align reads using more sensitive parameters for Illumina data:
./graphmap align -x illumina -r escherichia_coli.fa -d reads.fastq -o alignments.sam
# Load all reads in one batch and align only the first 1000 reads:
./graphmap align -B 0 -n 1000 -r escherichia_coli.fa -d reads.fastq -o alignments.sam
# Rebuild the index if it already exists:
./graphmap align --rebuild-index -r escherichia_coli.fa -d reads.fastq -o alignments.sam
# Generate only the index.
./graphmap align -I -r escherichia_coli.fa
# Run a debug version of GraphMap (build with "make debug") and verbose the SAM output to see various info about alignment:
./graphmap-debug align -b 3 -r escherichia_coli.fa -d reads.fastq -o alignments.sam
For additional information, help and bug reports please send an email to one of the following:
ivan.sovic@irb.hr, mile.sikic@fer.hr, nagarajann@gis.a-star.edu.sg
This work was supported by the IMaGIN platform (project No. 102 101 0025), through a grant from the Science and Engineering Research Council, funding to the Genome Institute of Singapore from the Agency for Science, Technology and Research (A*STAR), Singapore, and funding from the Croatian Science Foundation (Project no. UIP-11-2013-7353 - Algorithms for Genome Sequence Analysis).