readAligned {ShortRead} | R Documentation

## (Legacy) Read aligned reads and their quality scores into R representations

### Description

Import files containing aligned reads into an internal representation of the alignments, sequences, and quality scores. Most methods (see ‘details’ for exceptions) read all files into a single R object.

### Usage

    readAligned(dirPath, pattern=character(0), ...)

### Arguments

`dirPath` | A character vector (or other object; see methods defined on this generic) giving the directory path (relative or absolute; some methods also accept a character vector of file names) of aligned read files to be input.
`pattern` | The (`grep`-style) pattern describing file names to be read. The default (`character(0)`) results in (attempted) input of all files in the directory.
`...` | Additional arguments, used by methods. When `dirPath` is a character vector, the argument `type` must be provided. Possible values for `type` and their meaning are described below. Most methods implement `filter=srFilter()`, allowing objects of `SRFilter` to selectively returns aligned reads.

### Details

There is no standard aligned read file format; methods parse particular file types.

The `readAligned,character-method` interprets file types based on an additional `type` argument. Supported types are:

`type="SolexaExport"`

  This type parses `.*_export.txt` files following the documentation in the Solexa Genome Alignment software manual, version 0.3.0. These files consist of the following columns; consult Solexa documentation for precise descriptions. If parsed, values can be retrieved from `AlignedRead` as follows:

  Machine

    see below

  Run number

    stored in `alignData`

  Lane

    stored in `alignData`

  Tile

    stored in `alignData`

  X

    stored in `alignData`

  Y

    stored in `alignData`

  Multiplex index

    see below

  Paired read number

    see below

  Read

    `sread`

  Quality

    `quality`

  Match chromosome

    `chromosome`

  Match contig

    `alignData`

  Match position

    `position`

  Match strand

    `strand`

  Match description

    Ignored

  Single-read alignment score

    `alignQuality`

  Paired-read alignment score

    Ignored

  Partner chromosome

    Ignored

  Partner contig

    Ignored

  Partner offset

    Ignored

  Partner strand

    Ignored

  Filtering

    `alignData`

  The following optional arguments, set to `FALSE` by default, influence data input

  withMultiplexIndex

    When `TRUE`, include the multiplex index as a column `multiplexIndex` in `alignData`.

  withPairedReadNumber

    When `TRUE`, include the paired read number as a column `pairedReadNumber` in `alignData`.

  withId

    When `TRUE`, construct an identifier string as ‘Machine_Run:Lane:Tile:X:Y#multiplexIndex/pairedReadNumber’. The substrings ‘#multiplexIndex’ and ‘/pairedReadNumber’ are not present if `withMultiplexIndex=FALSE` or `withPairedReadNumber=FALSE`.

  withAll

    A convencience which, when `TRUE`, sets all `with*` values to `TRUE`.

  Note that not all paired read columns are interpreted. Different interfaces to reading alignment files are described in `SolexaPath` and `SolexaSet`.

`type="SolexaPrealign"`

  See SolexaRealign

`type="SolexaAlign"`

  See SolexaRealign

`type="SolexaRealign"`

  These types parse `s_L_TTTT_prealign.txt`, `s_L_TTTT_align.txt` or `s_L_TTTT_realign.txt` files produced by default and eland analyses. From the Solexa documentation, `align` corresponds to unfiltered first-pass alignments, `prealign` adjusts alignments for error rates (when available), `realign` filters alignments to exclude clusters failing to pass quality criteria.

  Because base quality scores are not stored with alignments, the object returned by `readAligned` scores all base qualities as `-32`.

  If parsed, values can be retrieved from `AlignedRead` as follows:

  Sequence

    stored in `sread`

  Best score

    stored in `alignQuality`

  Number of hits

    stored in `alignData`

  Target position

    stored in `position`

  Strand

    stored in `strand`

  Target sequence

    Ignored; parse using `readXStringColumns`

  Next best score

    stored in `alignData`

`type="SolexaResult"`

  This parses `s_L_eland_results.txt` files, an intermediate format that does not contain read or alignment quality scores.

  Because base quality scores are not stored with alignments, the object returned by `readAligned` scores all base qualities as `-32`.

  Columns of this file type can be retrieved from `AlignedRead` as follows (description of columns is from Table 19, Genome Analyzer Pipeline Software User Guide, Revision A, January 2008):

  Id

    Not parsed

  Sequence

    stored in `sread`

  Type of match code

    Stored in `alignData` as `matchCode`. Codes are (from the Eland manual): NM (no match); QC (no match due to quality control failure); RM (no match due to repeat masking); U0 (best match was unique and exact); U1 (best match was unique, with 1 mismatch); U2 (best match was unique, with 2 mismatches); R0 (multiple exact matches found); R1 (multiple 1 mismatch matches found, no exact matches); R2 (multiple 2 mismatch matches found, no exact or 1-mismatch matches).

  Number of exact matches

    stored in `alignData` as `nExactMatch`

  Number of 1-error mismatches

    stored in `alignData` as `nOneMismatch`

  Number of 2-error mismatches

    stored in `alignData` as `nTwoMismatch`

  Genome file of match

    stored in `chromosome`

  Position

    stored in `position`

  Strand

    (direction of match) stored in `strand`

  ‘N’ treatment

    stored in `alignData`, as `NCharacterTreatment`. ‘.’ indicates treatment of ‘N’ was not applicable; ‘D’ indicates treatment as deletion; ‘|’ indicates treatment as insertion

  Substitution error

    stored in `alignData` as `mismatchDetailOne` and `mismatchDetailTwo`. Present only for unique inexact matches at one or two positions. Position and type of first substitution error, e.g., 11A represents 11 matches with 12th base an A in reference but not read. The reference manual cited below lists only one field (`mismatchDetailOne`), but two are present in files seen in the wild.

`type="MAQMap", records=-1L`

  Parse binary `map` files produced by MAQ. See details in the next section. The `records` option determines how many lines are read; `-1L` (the default) means that all records are input. For `type="MAQMap"`, `dir` and `pattern` must match a single file.

`type="MAQMapShort", records=-1L`

  The same as `type="MAQMap"` but for map files made with Maq prior to version 0.7.0. (These files use a different maximum read length [64 instead of 128], and are hence incompatible with newer Maq map files.). For `type="MAQMapShort"`, `dir` and `pattern` must match a single file.

`type="MAQMapview"`

  Parse alignment files created by MAQ's ‘mapiew’ command. Interpretation of columns is based on the description in the MAQ manual, specifically

              ...each line consists of read name, chromosome, position,
              strand, insert size from the outer coordinates of a pair,
              paired flag, mapping quality, single-end mapping quality,
              alternative mapping quality, number of mismatches of the
              best hit, sum of qualities of mismatched bases of the best
              hit, number of 0-mismatch hits of the first 24bp, number
              of 1-mismatch hits of the first 24bp on the reference,
              length of the read, read sequence and its quality.

  The read name, read sequence, and quality are read as `XStringSet` objects. Chromosome and strand are read as `factor`s. Position is `numeric`, while mapping quality is `numeric`. These fields are mapped to their corresponding representation in `AlignedRead` objects.

  Number of mismatches of the best hit, sum of qualities of mismatched bases of the best hit, number of 0-mismatch hits of the first 24bp, number of 1-mismatch hits of the first 24bp are represented in the `AlignedRead` object as components of `alignData`.

  Remaining fields are currently ignored.

`type="Bowtie"`

  Parse alignment files created with the Bowtie alignment algorithm. Parsed columns can be retrieved from `AlignedRead` as follows:

  Identifier

    `id`

  Strand

    `strand`

  Chromosome

    `chromosome`

  Position

    `position`; see comment below

  Read

    `sread`; see comment below

  Read quality

    `quality`; see comments below

  Similar alignments

    `alignData`, ‘similar’ column; Bowtie v. 0.9.9.3 (12 May, 2009) documents this as the number of other instances where the same read aligns against the same reference characters as were aligned against in this alignment. Previous versions marked this as ‘Reserved’

  Alignment mismatch locations

    `alignData` ‘mismatch’, column

  NOTE: the default quality encoding changes to `FastqQuality` with ShortRead version 1.3.24.

  This method includes the argument `qualityType` to specify how quality scores are encoded. Bowtie quality scores are ‘Phred’-like by default, with `qualityType='FastqQuality'`, but can be specified as ‘Solexa’-like, with `qualityType='SFastqQuality'`.

  Bowtie outputs positions that are 0-offset from the left-most end of the `+` strand. `ShortRead` parses position information to be 1-offset from the left-most end of the `+` strand.

  Bowtie outputs reads aligned to the `-` strand as their reverse complement, and reverses the quality score string of these reads. `ShortRead` parses these to their original sequence and orientation.

`type="SOAP"`

  Parse alignment files created with the SOAP alignment algorithm. Parsed columns can be retrieved from `AlignedRead` as follows:

  id

    `id`

  seq

    `sread`; see comment below

  qual

    `quality`; see comment below

  number of hits

    `alignData`

  a/b

    `alignData` (`pairedEnd`)

  length

    `alignData` (`alignedLength`)

  +/-

    `strand`

  chr

    `chromosome`

  location

    `position`; see comment below

  types

    `alignData` (`typeOfHit`: integer portion; `hitDetail`: text portion)

  This method includes the argument `qualityType` to specify how quality scores are encoded. It is unclear from SOAP documentation what the quality score is; the default is ‘Solexa’-like, with `qualityType='SFastqQuality'`, but can be specified as ‘Phred’-like, with `qualityType='FastqQuality'`.

  SOAP outputs positions that are 1-offset from the left-most end of the `+` strand. `ShortRead` preserves this representation.

  SOAP reads aligned to the `-` strand are reported by SOAP as their reverse complement, with the quality string of these reads reversed. `ShortRead` parses these to their original sequence and orientation.

### Value

A single R object (e.g., `AlignedRead`) containing alignments, sequences and qualities of all files in `dirPath` matching `pattern`. There is no guarantee of order in which files are read.

### Author(s)

Martin Morgan <mtmorgan@fhcrc.org>, Simon Anders <anders@ebi.ac.uk> (MAQ map)

### See Also

The `AlignedRead` class.

Genome Analyzer Pipeline Software User Guide, Revision A, January 2008.

The MAQ reference manual, http://maq.sourceforge.net/maq-manpage.shtml#5, 3 May, 2008.

The Bowtie reference manual, http://bowtie-bio.sourceforge.net, 28 October, 2008.

The SOAP reference manual, http://soap.genomics.org.cn/soap1, 16 December, 2008.

### Examples

    sp <- SolexaPath(system.file("extdata", package="ShortRead"))
    ap <- analysisPath(sp)
    ## ELAND_EXTENDED
    (aln0 <- readAligned(ap, "s_2_export.txt", "SolexaExport"))
    ## PhageAlign
    (aln1 <- readAligned(ap, "s_5_.*_realign.txt", "SolexaRealign"))

    ## MAQ
    dirPath <- system.file('extdata', 'maq', package='ShortRead')
    list.files(dirPath)
    ## First line
    readLines(list.files(dirPath, full.names=TRUE)[[1]], 1)
    countLines(dirPath)
    ## two files collapse into one
    (aln2 <- readAligned(dirPath, type="MAQMapview"))

    ## select only chr1-5.fa, '+' strand
    filt <- compose(chromosomeFilter("chr[1-5].fa"),
                    strandFilter("+"))
    (aln3 <- readAligned(sp, "s_2_export.txt", filter=filt))

[Package ShortRead version 1.46.0 Index]