`RabbitTClust v.2.2.1`

RabbitTClust is a fast and memory-efficient genome clustering tool based on sketch-based distance estimations. It enables processing of large-scale datasets by combining dimensionality reduction techniques with streaming and parallelization on modern multi-core platforms. RabbitTClust supports classical single-linkage hierarchical (clust-mst) and greedy incremental clustering (clust-greedy) algorithms for different scenarios.

Installation

RabbitTClust v.2.2.1 can only support 64-bit Linux Systems.

The detailed update information for this version, as well as the version history, can be found in the version_history document.

Dependancy

cmake v.3.0 or later
c++14
zlib

Compile and install

git clone --recursive https://github.com/RabbitBio/RabbitTClust.git
cd RabbitTClust
./install.sh

Usage

# clust-mst, minimum-spanning-tree-based module for RabbitTClust
Usage: ./clust-mst [OPTIONS]
Options:
  -h,--help                   Print this help message and exit
  -t,--threads INT            set the thread number, default all CPUs of the platform
  -m,--min-length UINT        set the filter minimum length (minLen), genome length less than minLen will be ignore, default 10,000
  -c,--containment INT        use AAF distance with containment coefficient, set the containCompress, the sketch size is in proportion with 1/containCompress  -k,--kmer-size INT          set the kmer size
  -s,--sketch-size INT        set the sketch size for Jaccard Index and Mash distance, default 1000
  -l,--list                   input is genome list, one genome per line
  -e,--no-save                not save the intermediate files, such as sketches or MST
  -d,--threshold FLOAT        set the distance threshold for clustering
  -F,--function TEXT          set the sketch function, such as MinHash, KSSD, default MinHash
  -o,--output TEXT REQUIRED   set the output name of cluster result
  -i,--input TEXT Excludes: --append
                              set the input file, single FASTA genome file (without -l option) or genome list file (with -l option)
  --presketched TEXT          clustering by the pre-generated sketch files rather than genomes
  --premsted TEXT             clustering by the pre-generated mst files rather than genomes for clust-mst
  --newick-tree               output the newick tree format file for clust-mst
  --append TEXT Excludes: --input
                              append genome file or file list with the pre-generated sketch or MST files

# clust-greedy, greedy incremental clustering module for RabbitTClust
Usage: ./clust-greedy [OPTIONS]
Options:
  -h,--help                   Print this help message and exit
  -t,--threads INT            set the thread number, default all CPUs of the platform
  -m,--min-length UINT        set the filter minimum length (minLen), genome length less than minLen will be ignore, default 10,000
  -c,--containment INT        use AAF distance with containment coefficient, set the containCompress, the sketch size is in proportion with 1/containCompress  -k,--kmer-size INT          set the kmer size
  -s,--sketch-size INT        set the sketch size for Jaccard Index and Mash distance, default 1000
  -l,--list                   input is genome list, one genome per line
  -e,--no-save                not save the intermediate files, such as sketches or MST
  -d,--threshold FLOAT        set the distance threshold for clustering
  -F,--function TEXT          set the sketch function, such as MinHash, KSSD, default MinHash
  -o,--output TEXT REQUIRED   set the output name of cluster result
  -i,--input TEXT Excludes: --append
                              set the input file, single FASTA genome file (without -l option) or genome list file (with -l option)
  --presketched TEXT          clustering by the pre-generated sketch files rather than genomes
  --append TEXT Excludes: --input
                              append genome file or file list with the pre-generated sketch or MST files

Example:

# input is a file list, one genome path per line:
./clust-mst -l -i bact_refseq.list -o bact_refseq.mst.clust
./clust-greedy -l -i bact_genbank.list -o bact_genbank.greedy.clust

# input is a single genome file in FASTA format, one genome as a sequence:
./clust-mst -i bacteria.fna -o bacteria.mst.clust
./clust-greedy -i bacteria.fna -o bacteria.greedy.clust

# the sketch size (reciprocal of sampling proportion), kmer size, and distance threshold can be specified by -s (-c), -k, and -d options.
./clust-mst -l -k 21 -s 1000 -d 0.05 -i bact_refseq.list -o bact_refseq.mst.clust
./clust-greedy -l -k 21 -c 1000 -d 0.05 -i bact_genbank.list -o bact_genbank.greedy.clust


# for redundancy detection with clust-greedy, input is a genome file list:
# use -d to specify the distance threshold corresponding to various degrees of redundancy.
./clust-greedy -d 0.001 -l -i bacteria.list -o bacteria.out

# v.2.1.0 or later
# for last running of clust-mst, it generated a folder name in year_month_day_hour-minute-second format, such as 2023_05_06_08-49-15.
# this folder contains the sketch, mst files.
# for generator cluster from exist MST with a distance threshold of 0.045:
./clust-mst -d 0.045 --premsted 2023_05_06_08-49-15/ -o bact_refseq.mst.d.045.clust
# for generator cluster from exist sketches files of clust-mst with a distance threshold of 0.045:
./clust-mst -d 0.045 --presketched 2023_05_06_08-49-15/ -o bact_refseq.mst.d.045.clust

# for generator cluster from exist sketches of clust-greedy with a distance threshold of 0.001:
# folder 2023_05_06_08-49-15 contains the sketch files.
./clust-greedy -d 0.001 --presketched 2023_05_06_09-37-23/ -o bact_genbank.greedy.d.001.clust

# v.2.2.0 or later
# for generator cluster from exist part sketches (presketch_A_dir) and append genome set (genome_B.list) to incrementally clustering 
./clust-mst --presketched 2023_05_06_08-49-15/ -l --append genome_B.list -o append_refseq.mst.clust
./clust-mst --presketched 2023_05_06_09-37-23/ -l --append genome_B.list -o append_genbank.greedy.clust

# v.2.2.1 or later
# output the newick tree format for clust-mst, use the --newick-tree flag.
./clust-mst -l -i bacteria.list --newick-tree -o bacteria.mst.clust

Output

The output file is in a CD-HIT output format and is slightly different when running with or without -l input option.
When using the -l option, the input is expected to be a FASTA file list, with each file representing a genome. Without the -l option, the input should be a single FASTA file, with each sequence representing a genome.

Output format for a FASTA file list input

With -l* option, the tab-delimited values in the lines beginning with tab delimiters are:

local index in a cluster
global index of the genome
genome length
genome file name (including genome assembly accession number)
sequence name (first sequence in the genome file)
sequence comment (remaining part of the line)

Example:

the cluster 0 is:
    0   0   14782125nt  bacteria/GCF_000418325.1_ASM41832v1_genomic.fna     NC_021658.1     Sorangium cellulosum So0157-2, complete sequence
    1   1   14598830nt  bacteria/GCF_004135755.1_ASM413575v1_genomic.fna    NZ_CP012672.1   Sorangium cellulosum strain So ce836 chromosome, complete genome

the cluster 1 is:
    0   2   14557589nt  bacteria/GCF_002950945.1_ASM295094v1_genomic.fna    NZ_CP012673.1   Sorangium cellulosum strain So ce26 chromosome, complete genome

the cluster 2 is:
    0   3   13673866nt  bacteria/GCF_019396345.1_ASM1939634v1_genomic.fna   NZ_JAHKRM010000001.1    Nonomuraea guangzhouensis strain CGMCC 4.7101 NODE_1, whole genome shotgun sequence

......

Output format for a single FASTA file input

Without -l option, the tab-delimited values in the lines beginning with tab delimiters are:

local index in a cluster
global index of the genome
genome length
sequence name
sequence comment (remaining part of this line)

Example:

the cluster 0 is:
    0   0   11030030nt  NZ_GG657755.1   Streptomyces  himastatinicus ATCC 53653 supercont1.2, whole genome shotgun sequence
    1   1   11008137nt  NZ_RIBZ01000339.1   Streptomyces  sp. NEAU-LD23 C2041, whole genome shotgun sequence

the cluster 1 is:
    0   2   11006208nt  NZ_KL647031.1   Nonomuraea  candida strain NRRL B-24552 Doro1_scaffold1, whole genome shotgun sequence
    
the cluster 2 is:
    0   3   10940472nt  NZ_VTHK01000001.1   Amycolatopsis anabasis strain EGI 650086 RDPYD18112716_A.Scaf1, whole genome shotgun sequence

......

Output the newick tree format (v.2.2.1 or latter)

When the --newick-tree option is used, an additional output file will be generated in the Newick tree format with a suffix name of ".newick.tree".

Bug Report

We highly appreciate all bug reports, comments, and suggestions from our users.
Please feel free to raise any concerns or feedback with us without hesitation by issue.

Cite

Xu, X., Yin, Z., Yan, L. et al. RabbitTClust: enabling fast clustering analysis of millions of bacteria genomes with MinHash sketches. Genome Biol 24, 121 (2023). https://doi.org/10.1186/s13059-023-02961-6

XiaomingXu1995 / RabbitTClust