A fast constructor of the compressed de Bruijn graph from many genomes
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README.md

TwoPaCo 0.9.2

Release date: 18th May 2017

Authors

  • Ilia Minkin (Pennsylvania State University)
  • Son Pham (Salk Institute for Biological Studies)
  • Paul Medvedev (Pennsylvania State University)

Introduction

It is an implementation of the algorithm described in the paper "TwoPaCo: An efficient algorithm to build the compacted de Bruijn graph from many complete genomes".

This distribution contains two programs:

  • twopaco -- a tool for direct construction of the compressed graph from multiple complete genomes
  • graphdump -- a utility that turns output of twopaco into a text format

Test data

Links to the data used for bencharmking in the paper: https://github.com/medvedevgroup/TwoPaCo/blob/master/data.txt

Compilation

To compile the code, you need the following (Linux only):

  • CMake
  • A GCC compiler supporting C++11
  • Intel TBB library properly installed on your system. In other words, G++ should be able to find TBB libs

Once you've got all the things above, do the following:

  • Go to the root directory of the project and create the "build" folder
  • Go to the "build" directory
  • Run cmake ../src
  • Run make

This will make two targets: twopaco and graphdump. Compilation under other platforms is possible, portable makefiles are in progress.

TwoPaCo usage

To construct the graph (assuming you are in dir with "twopaco"), type:

./twopaco -f <filter_size> -k <value_of_k> <input_files>

This will constuct the compressed graph for the vertex size of <value_of_k> using 2^<filter_size> bits in the Bloom filter. The output file is a binary that can be either converted to a text file or read directly using an API (will be available soon).

Below is description of the other parameters.

K-mer size

This value sets the size of a vertex in the de Bruijn graph. Default is 25, to change, use:

-k <number> or --kvalue <number>

Note that:

  1. TwoPaCo uses k as the size of the vertex and (k + 1) as the size of the edge
  2. k must be odd

The maximum value of K supported by TwoPaCo is determined at the compile time. To increase the max value of K, increase the value "MAX_CAPACITY" defined in the header "vertexenumerator.h" and recompile. The value of "MAX_CAPACITY" should be at least (K + 4) / 32 + 1. Note that increasing the parameter will slow down the compilation.

Number of hash functions

The number of hash functions used for the Bloom filter. The default is five. To change, use:

-q <number> or --hashfnumber <number>

More hash functions increases the running time. At the same time, more hash functions may decrease the number of false positives and the memory usage.

Number of rounds

Number of computational rounds. Each rounds processses a separate subset of k-mers which reduces memory usage. The default is 1. To change, use:

-r <number> or --rounds <number>

The more the number of rounds, the longer the program works.

Number of threads

twopaco can be run with multiple threads. The default is 1. To change, use:

-t <number> or --threads <number>

Temporary directory

The directory for temporary files. The default is the current working directory. To change, use (the directory must exist):

--tmpdir <path_to_the_directory>

Output file name

The name of the output file. The default is "de_bruijn.bin". To change, use:

--o <file_name> or --outfile <file_name>

Running tests

If the flag is set, TwoPaCo will run a set of internal tests instead of processing the input file:

--test

The graphdump usage

This utility turns the binary file a text one. There are several output formats available. The folder "example" contains an example described in details.

GFF

In the next release I will add an option to output coordinates of all occurrences of the junctions in GFF format.

DOT

This format is used for visualization. The resulting DOT file can be converted into an image using Graphviz package:

http://www.graphviz.org/

To get the DOT file, use:

graphdump <twopaco_output_file> -f dot -k <value_of_k>

Note that the graph is a union of graphs built from both strands, with blue edges coming from the main strand and red ones from reverse one. The labels of the edges will indicate its position on a chromosome.

GFA

GFA is the most handy option. It explicitly represents the graph as a list of edges (non-branching paths in the non-compacted de Bruijn graph) graph and adjacencies between them. The file also contains all occurrences of the strings spelled by the paths in the input genomes.

In other words, it describes a colored de Bruijn graph where each path is mapped to several locations in the input ("colored"). TwoPaCo supports both GFA1 and GFA2. They are described here:

https://github.com/GFA-spec/GFA-spec

To get GFA output, run:

graphdummp <twopaco_output_file> -f gfa[version] -k <value_of_k> -s <input_genomes>

In the resulting file compacted non-branching paths are "segments" with "links" (GFA1) or "edges" (GFA2) containing them. "Containment" (GFA1) or "Fragment" (GFA2) records desrcibe the mapping between the non-branching paths in the graph and the input genomes. For GFA1, each input chromosome is also a "segment" described in the very beginning of the GFA file.

GFA1 only: each segment representing an input chromosome has the name of the corresponding header of the sequence in input FASTA file. In case if there are duplicate headers, one can add a prefix to segment names:

"s<number>_" + header of the sequence in input FASTA file

To do so, use the switch:

--prefix

For an example of GFA output and more detailed explanation, see the "example" folder.

Junctions List Format

In this format the output file only contains positions of junctions in the input genomes. As described in the paper, you can trivially restore information about edges from this junctions list. Note that junctions are mapped to genomes, i.e. one can reconstruct a colored graph from it. To get the junctions list, run:

graphdump -f seq -k <value_of_k>

This command will output a text file to the standard output. Each line will contain a triple indicating an occurence of junction:

<seq_id_i> <pos_i> <junction_id_i> 

The first number is the index number of the sequence, the second one is the position, and the third one is the junction id. The index number of the sequence is the order of the sequence in the input file(s). All positions/orders count from 0. Positions appear in the file in the same order they appear in the input genomes. The <junction_id> is a signed integer, the id of the junction that appears on the positive strand strand. A positive number indicates "direct" version of the junction, while a negative one shows the reverse complimentary version of the same junction. For example +1 and -1 are different versions of the same junction. This way, one can obtain all multi-edges of the graph with a linear scan, as described in the paper. For example, a sequence of of junctions ids:

a_1
a_2
a_3

Generates edges a_1 -> a_2, a_2 -> a_3 in the graph corresponding to the positive strand. To obtain the edges from the positive strand, one has to traverse them in the backwards order and negate signs, e.g. for the example above the sequence will be -a_3 -> -a_2 -> -a_1. One can also output junctions grouped by ids, it is useful for comparison between different graphs:

graphdump -f group -k <value_of_k>

In this format the i-th line line corresponds to the i-th junction and is of format:

<seq_id_0> <pos_0>; <seq_id_1> <pos_1>; ....

Where each pair "seq_id_i pos_j" corresponds to an occurence of the junction in sequence "seq_id_i" at position "pos_j". Sequence ids are just the numbers of sequences in the order they appear in the input. All positions count from 0.

Read The Binary File Directly

This is the most parsimonious option in terms of involved resources. One can read junctions and/or edges from the output file using a very simple C++ API. I will add the description in the future release. For now, one can use the sources of graphdump as a reference, it is relatively straightforward.

License

See LICENSE.txt

Contacts

Please e-mail your feedback at ivminkin@gmail.com.

You also can report bugs or suggest features using issue tracker at GitHub https://github.com/medvedevgroup/TwoPaCo

Citation

If you use TwoPaCo, please cite:

Ilia Minkin, Son Pham, and Paul Medvedev
"TwoPaCo: An efficient algorithm to build the compacted de Bruijn graph from many complete genomes"
Bioinformatics, 2016 doi:10.1093/bioinformatics/btw609

This project has been supported in part by NSF awards DBI-1356529, CCF-1439057, IIS-1453527, and IIS-1421908.