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PROGRAM:

NAME

dnaclust - program to cluster large number of short DNA sequences

SYNOPSIS

dnaclust {-i | --input} infile [{-s | --similarity} threshold]
[{-m | --multiple-alignment}] [{-d | --header}] [{-l | --left-gaps-allowed}]
[{-k | --k-mer-length} length] [{-a | --approximate-filter}] [--no-k-mer-filter]

dnaclust [{-h | --help} | {-v | --version}]

DESCRIPTION

This manual page documents briefly the dnaclust program.

dnaclust is a tool for clustering large number of short DNA sequences. The clusters are
created in such a way that the "radius" of each clusters is no more than the specified
threshold.

The input sequences to be clustered should be in Fasta format. The id of each sequence is
based on the first word of the seqeunce in the Fasta format. The first word is the prefix
of the header up to the first occurance of white space characters in the header. The
output is written to STDOUT. If you want the output to be written to a file, just redirect
the output (See Examples).

The output has two modes: the default clustering mode, and clustering with multiple
sequence alignment. In the clustering mode (without multiple alignment), each cluster will
be printed on a separate line. The line will contain the ids of the sequences in the
cluster. The first id in each line is the cluster center sequence id. Because of the way
our clusters are constructed, the length of the cluster center sequence is always greater
than or equal to the length of any of the sequences in the cluster. Please note that since
usually some clusters contain many sequences, the lines of the output may be very long. If
you want to visually inspect the output, please use the 'less -S', or an editor that does
not wrap long lines. The number of clusters can be found using 'wc -l'.

For more information about the multiple sequence alignment mode see the description of
--multiple-alignment option.

OPTIONS

The program follows the usual GNU command line syntax, with long options starting with two
dashes ('-'). A summary of options is included below.

--similarity threshold, -s threshold
The similarity threshold specifies the radius of the clusters created. This parameter
is a floating point number between 0 and 1. It is calculated based on semi-global
alignment of a sequence to the cluster center sequence. Namely similarity = 1 - (edit
distance) / (length of the shorter sequence). The edit distance is the minimum number
of insertions, deletions or substitutions necessary to aling a sequence to the cluster
center sequence. Our algorithms are faster when the similarity is higher.

--k-mer-length length, -k length
When you use the k-mer filter (which is enabled by default) you can specify the
maximum length of the k-mers used for filtering.

The longer k-mer lengths require more memory to store k-mer counts and the filtering
will be slower. However with the longer k-mer length, the filter will be more specific
and therefore the sequence alignment search may be faster.

There is a tradeoff between filtering and searching time. If you do not specify the
k-mer length a value of log4(median of the lengths of the input sequences) is picked
automatically. By using this option you can override the default value.

Keep in mind, however, that longer k-mer lengths would require more memory to store
the filtering data structures.

--approximate-filter , -a
By default the k-mer filter is 100 percent sensitive. This means that in the output
clustering, no two cluster centers are within the threshold distance from each other.
The exact filter, however, is somewhat slow. This option speeds up the filter by using
a heuristic. The use of the approximate filter may result in cluster centers that are
close, and a larger number clusters overall. However the approximate filter is usually
several times faster than the exact sensitive filter. Use this option if you are
clustering primarily to reduce the redundancy in the data, and do not care about the
quality of the clustering.

--allow-left-gaps , -l
With this option the distances are measured based on semi-global alignment. The
semi-global alignment allows for gaps without penalty on both ends of the shorter
sequence.

The default alignment is a one sided semi-global alignment. i.e. gaps are only allowed
at the right end of the shorter sequence without penalty. This behavior corresponds to
the data from targeted sequening of a region (e.g. of 16S ribosomal RNA gene).

--multiple-alignment, -m
Set the output format to show the multiple sequence alignment of each cluster. The
gaps in the alignments are represented with the dash '-' charachter.

The format of the MSA output is as follows: The MSA of each cluster spans several
lines. The MSA starts with a line containing charachter '#' followed by the number of
sequences in that cluster. The aligned sequences (which may contain gaps) follow in
the Fasta format. Each Fasta record will be composed of two lines. The header line and
the sequence line. Since each aligned sequence is output on a single line, the output
may contain very long lines. Please use 'less -S', or an editor that does not wrap
long lines for inspecting the MSA also.

--no-k-mer-filter
Disables the k-mer filter. Suitable for clustering very short sequences at a high
similarity threshold.

-d, --header
Write program options to output.

-h, --help
Show summary of options.

-v, --version
Show version of program.

EXAMPLES

./dnaclust file.fasta -l -s 0.98 -k 3 > clusters

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