# last-train

This script tries to find suitable score parameters (substitution and gap scores) for aligning some given sequences.

It (probabilistically) aligns the sequences using some initial score parameters, then estimates better score parameters based on the alignments, and repeats this procedure until the parameters stop changing.

The usage is like this:

    lastdb mydb reference.fasta
    last-train mydb queries.fasta

last-train prints a summary of each alignment step, followed by the final score parameters, in a format that can be read by lastal's -p option.

You can also pipe sequences into last-train, for example:

    zcat queries.fasta.gz | last-train mydb

## Options

> `-h, --help` | Show a help message, with default option values, and exit.

### Training options

> `--revsym` | Force the substitution scores to have reverse-complement symmetry, e.g. score(A→G) = score(T→C). This is often appropriate, if neither strand is "special".
> `--matsym` | Force the substitution scores to have directional symmetry, e.g. score(A→G) = score(G→A).
> `--gapsym` | Force the insertion costs to equal the deletion costs.
> `--pid=PID` | Ignore alignments with > PID% identity. This aims to optimize the parameters for low-similarity alignments, similarly to the BLOSUM matrices.
> `--sample-number=N` | Use N randomly-chosen chunks of the query sequences. The queries are chopped into fixed-length chunks (as if they were first concatenated into one long sequence). If there are ≤ N chunks, all are picked. Otherwise, if the final chunk is shorter, it is never picked. 0 means use everything.
> `--sample-length=L` | Use randomly-chosen chunks of length L.

All options below this point are passed to lastal to do the alignments: they are described in more detail at lastal.html.

### Initial parameter options

> `-r SCORE` | Initial match score.
> `-q COST` | Initial mismatch cost.
> `-p NAME` | Initial match/mismatch score matrix.
> `-a COST` | Initial gap existence cost.
> `-b COST` | Initial gap extension cost.
> `-A COST` | Initial insertion existence cost.
> `-B COST` | Initial insertion extension cost.

### Alignment options

> `-D LENGTH` | Query letters per random alignment. (See here.)
> `-E EG2` | Maximum expected alignments per square giga. (See here.)
> `-s NUMBER` | Which query strand to use: 0=reverse, 1=forward, 2=both.
> `-S NUMBER` | Score matrix applies to forward strand of: 0=reference, 1=query. This matters only if the scores lack reverse-complement symmetry.
> `-C COUNT` | Before extending gapped alignments, discard any gapless alignment whose query range lies in COUNT other gapless alignments with higher score-per-length. This aims to reduce run time.
> `-T NUMBER` | Type of alignment: 0=local, 1=overlap.
> `-m COUNT` | Maximum number of initial matches per query position.
> `-P COUNT` | Number of parallel threads.
> `-Q NUMBER` | Query sequence format: 0=fasta, 1=fastq-sanger. Important: if you use option -Q, last-train will only train the gap scores, and leave the substitution scores at their initial values.

## Bugs

- last-train assumes that gap lengths roughly follow a geometric distribution. If they do not (which is often the case), the results may be poor.

- last-train can fail for various reasons, e.g. if the sequences are too dissimilar. If it fails to find any alignments, you could try reducing the alignment significance threshold with option -D.