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\name{GAlignments-class}
\docType{class}
% Class:
\alias{class:GAlignments}
\alias{GAlignments-class}
% Constructor:
\alias{GAlignments}
\alias{updateObject,GAlignments-method}
% Accessors:
\alias{length,GAlignments-method}
\alias{names,GAlignments-method}
\alias{seqnames,GAlignments-method}
\alias{rname}
\alias{rname,GAlignments-method}
\alias{strand,GAlignments-method}
\alias{names<-,GAlignments-method}
\alias{seqnames<-,GAlignments-method}
\alias{rname<-}
\alias{rname<-,GAlignments-method}
\alias{strand<-,GAlignments-method}
\alias{cigar}
\alias{cigar,GAlignments-method}
\alias{qwidth}
\alias{qwidth,GAlignments-method}
\alias{start,GAlignments-method}
\alias{end,GAlignments-method}
\alias{width,GAlignments-method}
\alias{njunc}
\alias{njunc,GAlignments-method}
\alias{elementMetadata<-,GAlignments-method}
\alias{seqinfo,GAlignments-method}
\alias{seqinfo<-,GAlignments-method}
% Coercion:
\alias{grglist,GAlignments-method}
\alias{granges,GAlignments-method}
\alias{rglist,GAlignments-method}
\alias{ranges,GAlignments-method}
\alias{coerce,GAlignments,GRangesList-method}
\alias{coerce,GAlignments,GRanges-method}
\alias{coerce,GAlignments,RangesList-method}
\alias{coerce,GAlignments,Ranges-method}
\alias{as.data.frame,GAlignments-method}
\alias{coerce,GenomicRanges,GAlignments-method}
% Combining:
\alias{c,GAlignments-method}
% "show" method:
\alias{show,GAlignments-method}
% Other methods:
\alias{splitAsListReturnedClass,GAlignments-method}
% Old stuff:
\alias{GappedAlignments}
\alias{class:GappedAlignments}
\alias{GappedAlignments-class}
\alias{show,GappedAlignments-method}
\alias{ngap,GAlignments-method}
\title{GAlignments objects}
\description{
The GAlignments class is a simple container which purpose is
to store a set of genomic alignments that will hold just enough
information for supporting the operations described below.
}
\details{
A GAlignments object is a vector-like object where each element
describes a genomic alignment i.e. how a given sequence (called "query"
or "read", typically short) aligns to a reference sequence (typically
long).
Typically, a GAlignments object will be created by loading
records from a BAM (or SAM) file and each element in the resulting
object will correspond to a record. BAM/SAM records generally contain
a lot of information but only part of that information is loaded
in the GAlignments object. In particular, we discard the query
sequences (SEQ field), the query qualities (QUAL), the mapping qualities
(MAPQ) and any other information that is not needed in order to support
the operations or methods described below.
This means that multi-reads (i.e. reads with multiple hits in the
reference) won't receive any special treatment i.e. the various SAM/BAM
records corresponding to a multi-read will show up in the GAlignments
object as if they were coming from different/unrelated queries.
Also paired-end reads will be treated as single-end reads and the
pairing information will be lost (see \code{?\link{GAlignmentPairs}}
for how to handle aligned paired-end reads).
Each element of a GAlignments object consists of:
\itemize{
\item The name of the reference sequence. (This is the RNAME field
in a SAM/BAM record.)
\item The strand in the reference sequence to which the query is
aligned. (This information is stored in the FLAG field in a
SAM/BAM record.)
\item The CIGAR string in the "Extended CIGAR format" (see the SAM
Format Specifications for the details).
\item The 1-based leftmost position/coordinate of the clipped query
relative to the reference sequence. We will refer to it as
the "start" of the query. (This is the POS field in a SAM/BAM
record.)
\item The 1-based rightmost position/coordinate of the clipped query
relative to the reference sequence. We will refer to it as
the "end" of the query. (This is NOT explicitly stored in a
SAM/BAM record but can be inferred from the POS and CIGAR fields.)
Note that all positions/coordinates are always relative to
the first base at the 5' end of the plus strand of the reference
sequence, even when the query is aligned to the minus strand.
\item The genomic intervals between the "start" and "end" of the query
that are "covered" by the alignment. Saying that the full
[start,end] interval is covered is the same as saying that the
alignment contains no junction (no N in the CIGAR). It is then
considered to be a simple alignment. Note that a simple alignment
can have mismatches or deletions (in the reference). In other words,
a deletion (encoded with a D in the CIGAR) is NOT considered to
introduce a gap in the coverage, but a junction is.
}
Note that the last 2 items are not expicitly stored in the GAlignments
object: they are inferred on-the-fly from the CIGAR and the "start".
Optionally, a GAlignments object can have names (accessed thru the
\code{\link[base]{names}} generic function) which will be coming from
the QNAME field of the SAM/BAM records.
The rest of this man page will focus on describing how to:
\itemize{
\item Access the information stored in a GAlignments object
in a way that is independent from how the data are actually
stored internally.
\item How to create and manipulate a GAlignments object.
}
}
\section{Constructor}{
\describe{
\item{}{
\code{GAlignments(seqnames=Rle(factor()), pos=integer(0),
cigar=character(0),
strand=NULL, names=NULL, seqlengths=NULL, ...)}:
Low-level GAlignments constructor. Generally not used directly.
Named arguments in \code{...} are used as metadata columns.
}
}
}
\section{Accessors}{
In the code snippets below, \code{x} is a GAlignments object.
\describe{
\item{}{
\code{length(x)}:
Return the number of alignments in \code{x}.
}
\item{}{
\code{names(x)}, \code{names(x) <- value}:
Get or set the names on \code{x}.
See \code{\link{readGAlignments}} for how to automatically extract
and set the names when reading the alignments from a file.
}
\item{}{
\code{seqnames(x)}, \code{seqnames(x) <- value}:
Get or set the name of the reference sequence for each alignment
in \code{x} (see Details section above for more information about
the RNAME field of a SAM/BAM file).
\code{value} can be a factor, or a 'factor' \link[IRanges]{Rle},
or a character vector.
}
\item{}{
\code{rname(x)}, \code{rname(x) <- value}:
Same as \code{seqnames(x)} and \code{seqnames(x) <- value}.
}
\item{}{
\code{strand(x)}, \code{strand(x) <- value}:
Get or set the strand for each alignment in \code{x} (see Details
section above for more information about the strand of an alignment).
\code{value} can be a factor (with levels +, - and *), or a 'factor'
\link[IRanges]{Rle}, or a character vector.
}
\item{}{
\code{cigar(x)}:
Returns a character vector of length \code{length(x)}
containing the CIGAR string for each alignment.
}
\item{}{
\code{qwidth(x)}:
Returns an integer vector of length \code{length(x)}
containing the length of the query *after* hard clipping
(i.e. the length of the query sequence that is stored in
the corresponding SAM/BAM record).
}
\item{}{
\code{start(x)}, \code{end(x)}:
Returns an integer vector of length \code{length(x)}
containing the "start" and "end" (respectively) of the query
for each alignment. See Details section above for the exact
definitions of the "start" and "end" of a query.
Note that \code{start(x)} and \code{end(x)} are equivalent
to \code{start(granges(x))} and \code{end(granges(x))},
respectively (or, alternatively, to \code{min(rglist(x))} and
\code{max(rglist(x))}, respectively).
}
\item{}{
\code{width(x)}:
Equivalent to \code{width(granges(x))} (or, alternatively, to
\code{end(x) - start(x) + 1L}).
Note that this is generally different from \code{qwidth(x)}
except for alignments with a trivial CIGAR string (i.e. a
string of the form \code{"<n>M"} where <n> is a number).
}
\item{}{
\code{njunc(x)}:
Returns an integer vector of the same length as \code{x} containing
the number of junctions (i.e. N operations in the CIGAR) in each
alignment. Equivalent to \code{unname(elementLengths(rglist(x))) - 1L}.
}
\item{}{
\code{seqinfo(x)}, \code{seqinfo(x) <- value}:
Get or set the information about the underlying sequences.
\code{value} must be a \link{Seqinfo} object.
}
\item{}{
\code{seqlevels(x)}, \code{seqlevels(x) <- value}:
Get or set the sequence levels.
\code{seqlevels(x)} is equivalent to \code{seqlevels(seqinfo(x))}
or to \code{levels(seqnames(x))}, those 2 expressions being
guaranteed to return identical character vectors on a GAlignments
object. \code{value} must be a character vector with no NAs.
See \code{?\link{seqlevels}} for more information.
}
\item{}{
\code{seqlengths(x)}, \code{seqlengths(x) <- value}:
Get or set the sequence lengths.
\code{seqlengths(x)} is equivalent to \code{seqlengths(seqinfo(x))}.
\code{value} can be a named non-negative integer or numeric vector
eventually with NAs.
}
\item{}{
\code{isCircular(x)}, \code{isCircular(x) <- value}:
Get or set the circularity flags.
\code{isCircular(x)} is equivalent to \code{isCircular(seqinfo(x))}.
\code{value} must be a named logical vector eventually with NAs.
}
\item{}{
\code{genome(x)}, \code{genome(x) <- value}:
Get or set the genome identifier or assembly name for each sequence.
\code{genome(x)} is equivalent to \code{genome(seqinfo(x))}.
\code{value} must be a named character vector eventually with NAs.
}
\item{}{
\code{seqnameStyle(x)}:
Get or set the seqname style for \code{x}.
Note that this information is not stored in \code{x} but inferred
by looking up \code{seqnames(x)} against a seqname style database
stored in the \pkg{seqnames.db} metadata package (required).
\code{seqnameStyle(x)} is equivalent to \code{seqnameStyle(seqinfo(x))}
and can return more than 1 seqname style (with a warning)
in case the style cannot be determined unambiguously.
}
}
}
\section{Coercion}{
In the code snippets below, \code{x} is a GAlignments object.
\describe{
\item{}{
\code{grglist(x, use.mcols=FALSE,
order.as.in.query=FALSE,
drop.D.ranges=FALSE)},
\code{rglist(x, use.mcols=FALSE,
order.as.in.query=FALSE,
drop.D.ranges=FALSE)}:
Return either a \link{GRangesList} or a \link[IRanges]{RangesList}
object of length \code{length(x)} where the i-th element represents
the ranges (with respect to the reference) of the i-th alignment in
\code{x}.
More precisely, the \link[IRanges]{RangesList} object returned by
\code{rglist(x)} is a \link[IRanges]{CompressedIRangesList} object.
If \code{use.mcols} is TRUE and \code{x} has metadata columns on it
(accessible with \code{mcols(x)}), they're propagated to the returned
object.
The \code{order.as.in.query} toggle affects the order of the ranges
\emph{within} each top-level element of the returned object.
If \code{FALSE} (the default), then the ranges are ordered from 5' to 3'
in elements associated with the plus strand (i.e. corresponding to
alignments located on the plus strand), and from 3' to 5' in elements
associated with the minus strand. So, whatever the strand is, the ranges
are in ascending order (i.e. left-to-right).
If \code{TRUE}, then the order of the ranges in elements associated
with the \emph{minus} strand is reversed. So they end up being
ordered from 5' to 3' too, which means that they are now in decending
order (i.e. right-to-left). It also means that, when
\code{order.as.in.query=TRUE} is used, the ranges are
\emph{always} ordered consistently with the original "query template",
that is, in the order defined by walking the "query template" from the
beginning to the end.
If \code{drop.D.ranges} is \code{TRUE}, then deletions (D operations
in the CIGAR) are treated like junctions (N operations in the CIGAR),
that is, the ranges corresponding to deletions are dropped.
See Details section above for more information.
}
\item{}{
\code{granges(x, use.mcols=FALSE)}, \code{ranges(x)}:
Return either a \link{GRanges} or a \link[IRanges]{Ranges}
object of length \code{length(x)} where each element represents the
regions in the reference to which a query is aligned.
More precisely, the \link[IRanges]{Ranges} object returned by
\code{ranges(x)} is an \link[IRanges]{IRanges} object.
If \code{use.mcols} is TRUE and \code{x} has metadata columns on it
(accessible with \code{mcols(x)}), they're propagated to the returned
object.
}
\item{}{
\code{as(x, "GRangesList")}, \code{as(x, "GRanges")},
\code{as(x, "RangesList")}, \code{as(x, "Ranges")}:
Alternate ways of doing \code{grglist(x, use.mcols=TRUE)},
\code{granges(x, use.mcols=TRUE)}, \code{rglist(x, use.mcols=TRUE)},
and \code{ranges(x)}, respectively.
}
}
In the code snippet below, \code{x} is a \link[GenomicRanges]{GRanges}
object.
\describe{
\item{}{
\code{as(from, "GAlignments")}:
Creates a GAlignments object from a \link[GenomicRanges]{GRanges} object.
The metadata columns are propagated. cigar values are created from the
sequence width unless a "cigar" metadata column already exists in
\code{from}.
}
}
}
\section{Subsetting and related operations}{
In the code snippets below, \code{x} is a GAlignments object.
\describe{
\item{}{
\code{x[i]}:
Return a new GAlignments object made of the selected
alignments. \code{i} can be a numeric or logical vector.
}
}
}
\section{Combining}{
\describe{
\item{}{
\code{c(...)}:
Concatenates the GAlignments objects in \code{...}.
}
}
}
\section{Other methods}{
\describe{
\item{}{
\code{show(x)}:
By default the \code{show} method displays 5 head and 5 tail
elements. This can be changed by setting the global options
\code{showHeadLines} and \code{showTailLines}. If the object
length is less than (or equal to) the sum of these 2 options
plus 1, then the full object is displayed.
Note that these options also affect the display of \link{GRanges}
and \link{GAlignmentPairs} objects, as well as other objects
defined in the \pkg{IRanges} and \pkg{Biostrings} packages
(e.g. \link[IRanges]{Ranges} and \link[Biostrings]{DNAStringSet}
objects).
}
}
}
\references{
\url{http://samtools.sourceforge.net/}
}
\author{
H. Pages and P. Aboyoun
}
\seealso{
\itemize{
\item \code{\link{readGAlignments}} for reading genomic alignments
from a file (typically a BAM file) into a GAlignments object.
\item \link{GAlignmentPairs} objects for handling aligned paired-end
reads.
\item \link{junctions-methods} for extracting and summarizing junctions
from a GAlignments object.
\item \link[GenomicAlignments]{coverage-methods} for computing the
coverage of a GAlignments object.
\item \link[GenomicAlignments]{findOverlaps-methods} for finding
overlapping genomic alignments.
\item \code{\link[GenomicRanges]{seqinfo}} in the \pkg{GenomicRanges}
package for getting/setting/modifying the sequence information
stored in an object.
\item The \link[GenomicRanges]{GRanges} and
\link[GenomicRanges]{GRangesList} classes defined and documented
in the \pkg{GenomicRanges} package.
\item The \link[IRanges]{CompressedIRangesList} class defined and
documented in the \pkg{IRanges} package.
}
}
\examples{
library(Rsamtools) # for the ex1.bam file
ex1_file <- system.file("extdata", "ex1.bam", package="Rsamtools")
gal <- readGAlignments(ex1_file, param=ScanBamParam(what="flag"))
gal
## ---------------------------------------------------------------------
## A. BASIC MANIPULATION
## ---------------------------------------------------------------------
length(gal)
head(gal)
names(gal) # no names by default
seqnames(gal)
strand(gal)
head(cigar(gal))
head(qwidth(gal))
table(qwidth(gal))
head(start(gal))
head(end(gal))
head(width(gal))
head(njunc(gal))
seqlevels(gal)
## Rename the reference sequences:
seqlevels(gal) <- sub("seq", "chr", seqlevels(gal))
seqlevels(gal)
grglist(gal) # a GRangesList object
stopifnot(identical(unname(elementLengths(grglist(gal))), njunc(gal) + 1L))
granges(gal) # a GRanges object
rglist(gal) # a CompressedIRangesList object
stopifnot(identical(unname(elementLengths(rglist(gal))), njunc(gal) + 1L))
ranges(gal) # an IRanges object
## Modify the number of lines in 'show'
options(showHeadLines=3)
options(showTailLines=2)
gal
## Revert to default
options(showHeadLines=NULL)
options(showTailLines=NULL)
## ---------------------------------------------------------------------
## B. SUBSETTING
## ---------------------------------------------------------------------
gal[strand(gal) == "-"]
gal[grep("I", cigar(gal), fixed=TRUE)]
gal[grep("N", cigar(gal), fixed=TRUE)] # no junctions
## A confirmation that none of the alignments contains junctions (in
## other words, each alignment can be represented by a single genomic
## range on the reference):
stopifnot(all(njunc(gal) == 0))
## Different ways to subset:
gal[6] # a GAlignments object of length 1
grglist(gal)[[6]] # a GRanges object of length 1
rglist(gal)[[6]] # a NormalIRanges object of length 1
## Unlike N operations, D operations don't introduce gaps:
ii <- grep("D", cigar(gal), fixed=TRUE)
gal[ii]
njunc(gal[ii])
grglist(gal[ii])
## qwidth() vs width():
gal[qwidth(gal) != width(gal)]
## This MUST return an empty object:
gal[cigar(gal) == "35M" & qwidth(gal) != 35]
## but this doesn't have too:
gal[cigar(gal) != "35M" & qwidth(gal) == 35]
}
\keyword{methods}
\keyword{classes}
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