Discontiguous Mega BLAST

# What is discontiguous Mega BLAST?

    This version of Mega BLAST is designed specifically for comparison of diverged
    sequences, especially sequences from different organisms, which have alignments
    with low degree of identity, where the original Mega BLAST is not very
    effective. The major difference is in the use of the 'discontiguous word'
    approach to finding initial offset pairs, from which the gapped extension is
    then performed.

    Both Mega BLAST and all previous versions of nucleotide-nucleotide BLAST look
    for exact matches of certain length as the starting points for gapped
    alignments. When comparing less conserved sequences, i.e. when the expected
    share of identity between them is e.g. 80% and below, this traditional approach
    becomes much less productive than for the higher degree of conservation.
    Depending on the length of the exact match to start the alignments from, it
    either misses a lot of statistically significant alignments, or on the contrary
    finds too many short random alignments.

    According to [1], as well as our own probability simulations, it turns out that
    if initial 'words' are based not on the exact match, but on a match of a certain
    set of nonconsecutive positions within longer segments of the sequences, the
    productivity of the word finding algorithm is much higher. This way fewer words
    are found overall, but more of them end up producing statistically significant
    alignments, than in the case of contiguous words of the same, and even shorter
    length than the number of matched positions in the discontiguous word.

    As an example, we can define a pattern (template) of 0s and 1s of length e.g.
    21:
    100101100101100101101. For each pair of offsets in the query and subject
    sequences that are being compared, we compare the 21 nucleotide segments in
    these sequences ending at these offsets, and require only those positions in
    those segments to match that correspond to the 1s in the above template.

    There are several advantages in using this approach. First, the conditional
    probabilities of finding word hits satisfying discontiguous templates given the
    expected identity percentage in the alignments between two sequences, are higher
    than for contiguous words with the same number of positions required matched.
    If two word hits are required to initiate a gapped extension, the effect
    of the discontiguous word approach is even larger. In both cases higher
    sensitivity is achieved because there is less correlation between successive
    words as the database sequence is scanned across the query sequence.
    Second, when comparing coding sequences, the conservation of the
    third nucleotides in every codon is not essential, so there is no need to
    require it when matching initial words. This implies the advantage of using
    templates based on the '110' pattern, which are called 'coding'.
    Finally, to achieve even higher sensitivity, one might combine two different
    discontiguous word templates and require any one of them to match at a given
    position to qualify it for the initial word hit.

    The following options specific to this approach are supported:

    Template length: 16, 18, 21.
    Word size (i.e. number of 1s in the template): 11, 12
    Template type: coding, non-coding.
    Require two words for extension: yes/no.

    The 'coding' templates are based on the 110 pattern, although more 0s are
    required for most of them, so some of the patterns become 010 or 100. These are
    the most effective for comparison of coding regions.

    The non-coding templates attempt to minimize the correlation between successive
    words, when the database sequence is shifted by 4 positions against the query
    sequence. This means more 1s are concentrated at the ends of the template (at
    least 3 on each side).

    When the option to require two words for extension is chosen, two word hits
    matching the template must be found within a distance of 50 nucleotides of one
    another.

    Below are the exact discontiguous word template patterns for different combinations
    of word sizes and lengths:

       W = 11, t = 16, coding:     1101101101101101
       W = 11, t = 16, non-coding: 1110010110110111
       W = 12, t = 16, coding:     1111101101101101
       W = 12, t = 16, non-coding: 1110110110110111
       W = 11, t = 18, coding:     101101100101101101
       W = 11, t = 18, non-coding: 111010010110010111
       W = 12, t = 18, coding:     101101101101101101
       W = 12, t = 18, non-coding: 111010110010110111
       W = 11, t = 21, coding:     100101100101100101101
       W = 11, t = 21, non-coding: 111010010100010010111
       W = 12, t = 21, coding:     100101101101100101101
       W = 12, t = 21, non-coding: 111010010110010010111

    [1] Ma, B., Tromp, J., Li, M., "PatternHunter: faster and more sensitive
    homology search", Bioinformatics 2002 Mar;18(3):440-5