%\VignetteIndexEntry{Introduction to GenomicFiles}
%\VignetteDepends{GenomicAlignments, RNAseqData.HNRNPC.bam.chr14}
%\VignetteKeywords{parallel, sequencing, fileIO}
%\VignettePackage{GenomicFiles}

\documentclass{article}

<<style, eval=TRUE, echo=FALSE, results=tex>>=
BiocStyle::latex()
@

\title{Introduction to \Biocpkg{GenomicFiles}}
\author{Valerie Obenchain, Michael Love, Martin Morgan}
\date{Last modified: October 2014; Compiled: \today}

\begin{document}

\maketitle

\tableofcontents

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\section{Introduction}
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This vignette illustrates how to use the \Biocpkg{GenomicFiles} package for
distributed computing across files. The functions in \Rcode{GenomicFiles} 
manipulate and combine data subsets via two user-supplied functions, MAP and 
REDUCE. These are similar in spirit to \Rcode{Map} and \Rcode{Reduce} in 
\Rpackage{base} \R{}. Together they provide a flexible interface to extract, 
manipulate and combine data. Both functions are executed in the distributed
step which means results are combined on a single worker, not across workers.

We assume the reader has some previous experience with \R{} and with basic 
manipulation of ranges objects such as \Rcode{GRanges} and \Rcode{GAlignments} 
and file classes such as \Rcode{BamFile} and \Rcode{BigWigFile}. See the 
vignettes and documentation in \Biocpkg{GenomicRanges}, 
\Biocpkg{GenomicAlignments}, \Biocpkg{Rsamtools} and \Biocpkg{rtracklayer} for 
an introduction to these classes.

The \Rpackage{GenomicFiles} package is available at bioconductor.org
and can be downloaded via \Rcode{BiocManager::install}:

<<install, eval=FALSE>>=
if (!require("BiocManager"))
    install.packages("BiocManager")
BiocManager::install("GenomicFiles")
@

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\section{Quick Start}
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\Rpackage{GenomicFiles} offers functions for the parallel extraction 
and combination of data subsets. A user-defined MAP function extracts and 
manipulates data while an optional REDUCE function consolidates 
the output of MAP. 

<<quick_start-load, results=hide>>=
library(GenomicFiles)
@

Ranges can be a \Rcode{GRanges}, \Rcode{GRangesList} or \Rcode{GenomicFiles}
class.
<<quick_start-ranges>>=
gr <- GRanges("chr14", IRanges(c(19411500 + (1:5)*20), width=10))
@

File are supplied as a character vector or list of *File classes such as
\Rcode{BamFile}, \Rcode{BigWigFile} etc. 
<<class-bam-data>>=
library(RNAseqData.HNRNPC.bam.chr14)
fls <- RNAseqData.HNRNPC.bam.chr14_BAMFILES
@

The MAP function extracts and manipulates data subsets. Here we compute 
pileups for a given range and file.
<<quick_start-MAP>>=
MAP <- function(range, file, ...) {
    requireNamespace("Rsamtools")
    Rsamtools::pileup(file, scanBamParam=Rsamtools::ScanBamParam(which=range))
}
@

\Rcode{reduceByFile} sends each file to a worker where MAP is applied to each
file / range combination. When \Rcode{summarize=TRUE} the output is
a \Rcode{SummarizedExperiment} object.
<<quick_start-reduceByFile>>=
se <- reduceByFile(gr, fls, MAP, summarize=TRUE)
se
@

Results are stored in the \Rcode{assays} slot.
<<quick_start-assays>>=
dim(assays(se)$data)  ## ranges x files
@

\Rcode{reduceByRange} sends each range to a worker and extracts the same range 
from all files. Adding a reducer to this example combines the pileups
from each range across files.
<<quick_start-MAP-REDUCE-reduceByRange>>=
REDUCE <- function(mapped, ...) {
    cmb = do.call(rbind, mapped)
    xtabs(count ~ pos + nucleotide, cmb)
}

lst <- reduceByRange(gr, fls, MAP, REDUCE, iterate=FALSE)
@

The result is a list where each element is a summary table of counts for a
single range across all 8 files.
<<quick_start-result>>=
head(lst[[1]], 3)
@

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\section{Overview of classes and functions}
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\subsection{\Rcode{GenomicFiles} class}

The \Rcode{GenomicFiles} class is a matrix-like container where rows represent
ranges of interest and columns represent files. The object can be subset on
files and / or ranges to perform different experimental runs. The class 
inherits from \Rcode{RangedSummarizedExperiment} but does not (as of yet)
make use of the \Rcode{elementMetadata} and \Rcode{assays} slots.

<<overview-GenomicFiles>>=
GenomicFiles(gr, fls)
@

A \Rcode{GenomicFiles} can be used as the \Rcode{ranges} argument to the
functions in this package. When \Rcode{summarize=TRUE}, data from the common
slots are transferred to the \Rcode{SummarizedExperiment} result.  NOTE: Results
can only be put into a \Rcode{SummarizedExperiment} when no reduction is
performed because of the matching dimensions requirement (i.e., a REDUCE
collapses the results in one dimension).

\subsection{Functions}

Functions in \Rcode{GenomicFiles} manipulate and combine data across or within
files using the parallel infrastructure provided in \Rcode{BiocParallel}.
Files and ranges are sent to workers along with MAP and REDUCE functions. The
MAP extracts and/or manipulates data and REDUCE consolidates the results from
MAP.  Both MAP and REDUCE are executed in the distributed step and therefore
reduction occurs on data from the same worker, not across workers.

The chart in Figure \ref{reduceByRange_flow} represents the division of labor
in \Rcode{reduceByRange} and \Rcode{reduceRanges} with 3 files and 4 ranges.
These functions split the problem by range which allows subsets (i.e., the same
range) to be combined across different files.  \Rcode{reduceByRange} iterates
through the files, invoking MAP and REDUCE for each range / file combination.
This approach allows ranges extracted from the files to be kept separate or
combined before the next call to \Rcode{MAP} based on whether or not a
\Rcode{REDUCE} is supplied.

\Rcode{reduceRanges} applies \Rcode{MAP} to each range / file combination and 
REDUCEs the output of all MAP calls. \Rcode{REDUCE} usually plays a minor 
role by concatenating or unlisting results.


\begin{figure}[!h]
  \begin{center}
    \includegraphics{reduceByRange_flow.png}
    \caption{Mechanics of \Rcode{reduceByRange} and \Rcode{reduceRanges}}
    \label{reduceByRange_flow}
  \end{center}
\end{figure}

In contrast to the `byRange` approach, \Rcode{reduceByFile} and
\Rcode{reduceFiles} (Figure \ref{reduceByFile_flow}) split the problem by file.
Files are sent to different workers with the set of ranges
allowing subsets (i.e., multiple ranges) from the same file to be combined.
\Rcode{reduceByFile} invokes \Rcode{MAP} for each file / range combination 
allowing potential \Rcode{REDUCE} after each MAP step.

\Rcode{reduceFiles} applies \Rcode{MAP} to each range / file combination and
REDUCEs the output of all MAP calls. \Rcode{REDUCE} usually plays a minor 
role by concatenating or unlisting results.

\begin{figure}[!h]
  \begin{center}
    \includegraphics{reduceByFile_flow.png}
    \caption{Mechanics of \Rcode{reduceByFile} and \Rcode{reduceFiles}}
    \label{reduceByFile_flow}
  \end{center}
\end{figure}

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\section{Queries across files: \Rcode{reduceByRange} and \Rcode{reduceRanges}}
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
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The \Rcode{reduceByRange} and \Rcode{reduceRanges} functions are designed for
analyses that compare or combine data subsets across files. The first example in
this section computes pileups on subsets from individual files then sums over
all files.  The second example computes coverage on a group of ranges for each
file then performs a basepair-level $t$-test across files. The $t$-test example
also demonstrates how to use a blocking factor to differentiate files by
experimental group (e.g., case vs control).
\pagebreak

\subsection{Pileup summaries}

In this example nucleotide counts (pileups) are computed for the same ranges
in each file (MAP step). Pileups are then summed by position resulting in a 
single table for each range across all files (REDUCE step).

Create a \Rclass{GRanges} with regions of interest:
<<pileups-ranges>>=
gr <- GRanges("chr14", IRanges(c(19411677, 19659063, 105421963, 
              105613740), width=20))
@

The \Rcode{bam2R} function from the \Rpackage{deepSNV} package
is used to compute the statistics. The MAP invokes \Rcode{bam2R} 
and retains only the nucleotide counts (see ?\Rcode{bam2R} for other 
output fields). Counts from the reference strand are uppercase and counts 
from the complement are lowercase.

Because the \Rcode{bam2R} function is not explicitly passed through the
MAP, \Rcode{deepSNV} must be loaded on each worker so the
function can be found.

<<pileups-MAP>>=
MAP <- function(range, file, ...) {
    requireNamespace("deepSNV")
    ct = deepSNV::bam2R(file, 
                        GenomeInfoDb::seqlevels(range), 
                        GenomicRanges::start(range), 
                        GenomicRanges::end(range), q=0)
    ct[, c("A", "T", "C", "G", "a", "t", "c", "g")]
}
@

With no REDUCE function, the output is a list the same length as the number
of ranges where each list element is the length of the number of files.
\begin{verbatim}
pile1 <- reduceByRange(gr, fls, MAP) 
> length(pile1)
[1] 4
> elementNROWS(pile1)
[1] 8 8 8 8
\end{verbatim}

Next add a REDUCE to sum the counts by position.
<<pileups-REDUCE>>=
REDUCE <- function(mapped, ...)
    Reduce("+", mapped)
@
 
The output is again a list with the same length as the number of ranges but the
element lengths have been reduced to 1.
<<pileups-reduceByRange>>=
pile2 <- reduceByRange(gr, fls, MAP, REDUCE) 
length(pile2)
elementNROWS(pile2)
@

Each element is a matrix of counts (position by nucleotide) for a single range
summed over all files.
<<pileups-res>>=
head(pile2[[1]])
@

\subsection{Basepair-level $t$-test with case / control groups}

In this example coverage is computed for a region of interest in multiple files.
A grouping variable that defines case / control status is passed as an extra
argument to \Rcode{reduceByRange} and used in the reduction step to perform
the $t$-test.

Define ranges of interest,
<<ttest-ranges>>=
roi <- GRanges("chr14", IRanges(c(19411677, 19659063, 105421963,
               105613740), width=20))
@

and assign the case, control grouping of files. (Grouping is arbitrary in this
example.)
<<ttest-group>>=
grp <- factor(rep(c("A","B"), each=length(fls)/2))
@ 

The MAP reads in alignments from each BAM file and computes coverage. Coverage 
is coerced from an RleList to numeric vector for later use in the $t$-test.
<<ttest-MAP>>=
MAP <- function(range, file, ...) {
    requireNamespace("GenomicAlignments")
    param <- Rsamtools::ScanBamParam(which=range)
    as.numeric(unlist(
        GenomicAlignments::coverage(file, param=param)[range], use.names=FALSE))
}
@ 

REDUCE combines the coverage vectors into a matrix, identifies all-zero rows,
and performs row-wise $t$-testing using the \Rcode{rowttests} function from the 
\Biocpkg{genefilter} package. The index of which rows correspond to which 
basepair of the original range is stored as a column \Robject{offset}.
<<ttest-REDUCE>>=
REDUCE <- function(mapped, ..., grp) {
    mat = simplify2array(mapped)
    idx = which(rowSums(mat) != 0)
    df = genefilter::rowttests(mat[idx,], grp)
    cbind(offset = idx - 1, df)
}
@ 

The file grouping is passed as an extra argument to \Rcode{reduceByRange}.
\Rcode{iterate=FALSE} postpones the reduction until coverage vectors for all 
files have been computed. This delay is necessary because REDUCE uses the file 
grouping factor to perform the $t$-test and relies on the coverage vectors for
all files to be present. 
<<ttest-results, eval=FALSE>>=
ttest <- reduceByRange(roi, fls, MAP, REDUCE, iterate=FALSE, grp=grp)
@

The result is a list of summary tables of basepair-level $t$-test statistics
for each range across all files.
\begin{verbatim}
> head(ttest[[1]], 3)
  offset statistic   dm   p.value
1      0 1.1489125 2.75 0.2943227
2      1 0.9761871 2.25 0.3666718
3      2 0.8320503 1.50 0.4372365
\end{verbatim}

These tables can be added to the \Rcode{roi} GRanges as a metadata column.
\begin{verbatim}
mcols(roi)$ttest <- ttest 
> head(roi)
GRanges object with 4 ranges and 1 metadata column:
      seqnames                 ranges strand |    ttest
         <Rle>              <IRanges>  <Rle> |   <list>
  [1]    chr14 [ 19411677,  19411696]      * | ########
  [2]    chr14 [ 19659063,  19659082]      * | ########
  [3]    chr14 [105421963, 105421982]      * | ########
  [4]    chr14 [105613740, 105613759]      * | ########
  -------
  seqinfo: 1 sequence from an unspecified genome; no seqlengths
\end{verbatim}

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\section{Queries within files: \Rcode{reduceByFile} and \Rcode{reduceFiles}}
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\Rcode{reduceByFile} and \Rcode{reduceFiles} compare or combine data subsets
within files. \Rcode{reduceByFile} allows for more fine-tuned manipulation over
the subset for each range / file combination. If differentiating between ranges
is not important, \Rcode{reduceFiles} can be used to treat the ranges as a
group.

In this section read junctions are counted for individual subsets within a file
then combined based on user-defined selection criteria. Another example computes
coverage over complete BAM files by streaming over a set of continuous ranges.
The coverage example is performed with both \Rcode{reduceByFile} and
\Rcode{reduceFiles} to demonstrate the passing ranges to MAP individually vs all
at once. The last example uses a MAP function to chunk through subsets when the 
data are too large for available memory.

\subsection{Counting read junctions}

This example highlights how \Rcode{reduceByFile} allows detailed control
over the combination of data subsets from distinct ranges within the same 
file.

Define ranges of interest.
<<junctions-ranges>>=
gr <- GRanges("chr14", IRanges(c(19100000, 106000000), width=1e7))
@

The MAP produces a table of junction counts (i.e., 'N' operations in the 
CIGAR) for each range.
<<junctions-MAP>>=
MAP <- function(range, file, ...) {
    requireNamespace("GenomicAlignments") ## for readGAlignments()
                                          ## ScanBamParam()
    param = Rsamtools::ScanBamParam(which=range)
    gal = GenomicAlignments::readGAlignments(file, param=param)
    table(GenomicAlignments::njunc(gal))
} 
@

Create a GenomicFiles object.
<<junctions-GenomicFiles>>=
gf <- GenomicFiles(gr, fls)
gf
@

The GenomicFiles object or any subset of the object can be used as the
\Rcode{ranges} argument to functions in \Rcode{GenomicFiles}.
Here the object is subset on 3 files and both ranges.
<<junctions-counts1>>=
counts1 <- reduceByFile(gf[,1:3], MAP=MAP)
length(counts1)        ## 3 files
elementNROWS(counts1)  ## 2 ranges
@

Each list element has a table of counts for each range.
<<junctions-counts1-show>>=
counts1[[1]]
@

Add a reducer that combines counts for records in each range with exactly 
1 junction.
<<junctions-REDUCE>>=
REDUCE <- function(mapped, ...)
    sum(sapply(mapped, "[", "1"))

reduceByFile(gr, fls, MAP, REDUCE)
@

Next invoke \Rcode{reduceFiles} with the same files and MAP function.
\Rcode{reduceFiles} treats all ranges as a group and counts junctions for
all ranges simultaneously. 
<<junctions-counts2>>=
counts2 <- reduceFiles(gf[,1:3], MAP=MAP)
@

In the \Rcode{reduceByFile} example junctions were counted for each range
individually which allowed us to see results for the individual ranges and
combine them on the fly based on specific criteria. In contrast,
\Rcode{reduceFiles} counts junctions for all ranges simultaneously. 
<<junctions-counts2-show>>=
## reduceFiles returns counts for all ranges.
counts2[[1]]
## reduceByFile returns counts for each range separately.
counts1[[1]]
@

\subsection{Coverage 1: \Rcode{reduceByFile}}

Files that are too large to fit in memory can be streamed over by creating `tiles` or
ranges that span the whole file. The \Rcode{tileGenome} function creates a set
of continuous ranges that span a given seqlength(s). The sample BAM files
contain only chr14 so we extract the appropriate seqlength from the BAM files
and use it in \Rcode{tileGenome}. In this example we create 5 ranges but the
optimal value for \Rcode{ntile} will depend on the application and the size of
the chromosome (or genome) to be tiled.
<<coverage1-tiles>>=
chr14_seqlen <- seqlengths(seqinfo(BamFileList(fls))["chr14"])
tiles <- tileGenome(chr14_seqlen, ntile=5)
@

\Rcode{tiles} is a GRangesList of length \Rcode{ntile} with one range
per element.
<<coverage1-tiles-show>>=
tiles
@

MAP computes coverage for each range. The sum of coverage across all positions
is recorded along with the width of the range.
<<coverage1-MAP>>=
MAP = function(range, file, ...) {
    requireNamespace("GenomicAlignments")  ## for ScanBamParam() and coverage()
    param = Rsamtools::ScanBamParam(which=range)
    rle = GenomicAlignments::coverage(file, param=param)[range]
    c(width = GenomicRanges::width(range), 
      sum = sum(S4Vectors::runLength(rle) * S4Vectors::runValue(rle))) 
}
@

REDUCE sums the width and coverage for all ranges in `tiles`. 
<<coverage1-REDUCE>>=
REDUCE = function(mapped, ...) {
    Reduce(function(i, j) Map("+", i, j), mapped)
}
@

When \Rcode{iterate=TRUE} REDUCE is applied after each MAP step. Iterating
prevents the data from growing too large on the worker. The total width and
coverage sum for all ranges are returned for each file.
<<coverage1-results, eval=FALSE>>=
cvg1 <- reduceByFile(tiles, fls, MAP, REDUCE, iterate=TRUE)
@
\begin{verbatim}
  > cvg1[1]
  $ERR127306
  $ERR127306$width
  [1] 107349540

  $ERR127306$sum.chr14
  [1] 57633506
\end{verbatim}

\subsection{Coverage 2: \Rcode{reduceFiles}}

In the first coverage example we used \Rcode{reduceByFile} to invoke MAP for
each file / range combination. This approach is useful when analyses
require data manipulation at the level of each file / range subset prior to
reduction. For many applications, however, distinguishing between ranges is not
important and the overhead of an lapply over all ranges may be costly.

An alternative is to use \Rcode{reduceFiles} which passes all ranges as a single
argument to MAP. The ranges can be used to create a `param` or passed as an 
argument to another function that operates on multiple ranges at
at time.


This MAP computes coverage on all ranges at once and returns an RleList.
<<coverage2-MAP>>=
MAP = function(range, file, ...) {
    requireNamespace("GenomicAlignments")  ## for ScanBamParam() and coverage()
    GenomicAlignments::coverage(
        file,
        param=Rsamtools::ScanBamParam(which=range))[range]
}
@

REDUCE extracts the RleList from `mapped` and collapses the coverage. Note that
reduction could have be done in the MAP step on the output of coverage.  Because
all ranges are passed as a single argument, MAP is only called once on each
worker. Consequences of a single invocation are (1) reduction can be done at the
end of the MAP or by REDUCE and (2) REDUCE cannot be applied iteratively (this
requires more than a single output from MAP).
<<coverage2-REDUCE>>=
REDUCE = function(mapped, ...) {
    sapply(mapped, Reduce, f = "+")
}
@

Recall `tiles` is a GRangesList with one range per list element. We have no
need for the grouping in this example so we pass `tiles` as a GRanges.
<<coverage2-results>>=
cvg2 <- reduceFiles(unlist(tiles), fls, MAP, REDUCE)
@

Output is a list of length 8 where each element is a single Rle of coverage 
for all ranges.
<<coverage2-show>>=
cvg2[1]
@

\subsection{Coverage 3: \Rcode{reduceFiles} with chunking}

Continuing with the same coverage example. Now let's assume the result from
calling \Rcode{coverage} with all ranges in `tiles` does not fit in available
memory. We need a way to chunk through the ranges.

One option is to use \Rcode{reduceByFile} to lapply through each range in
`tiles` individually and then apply a reducer as we did in the first coverage
example. Because the `tiles` GRangesList has only one range per list element
this approach may be inefficient for a large number of ranges. To reduce the
number of iterations in the lapply, the ranges in `tiles` could be re-grouped
into a GRangesList with more than one range per element.

Another approach is to write your own MAP function that chunks through the
ranges. This has the advantage that, if resources are available, an additional
level of parallel dispatch can be implemented.

MAP creates an index over the ranges which are passed to \Rcode{bplapply}.
The data are subset on each worker, coverage is computed and reduced for
the ranges in the chunk.
<<coverage3-MAP>>=
MAP = function(range, file, ...) {
    requireNamespace("BiocParallel")  ## for bplapply()
    nranges = 2
    idx = split(seq_along(range), ceiling(seq_along(range)/nranges))
    BiocParallel::bplapply(idx, 
        function(i, range, file) {
            requireNamespace("GenomicAlignments")  ## ScanBamParam(), coverage()
            chunk = range[i]
            param = Rsamtools::ScanBamParam(which=chunk)
            cvg = GenomicAlignments::coverage(file, param=param)[chunk]
            Reduce("+", cvg)            ## collapse coverage within chunks
        }, range, file)
}
@

REDUCE extracts and collapses the RleList of coverage for all chunks.
<<coverage3-REDUCE>>=
REDUCE = function(mapped, ...) {
    sapply(mapped, Reduce, f = "+")
}
@

Again `tiles` are passed as a GRanges so the chunking in MAP defines the 
groups, not the structure of the GRangesList. Output is a list of length 8 
where each list element is a single Rle of coverage. 
<<coverage3-results, eval=FALSE>>=
cvg3 <- reduceFiles(unlist(tiles), fls, MAP, REDUCE)
@
\begin{verbatim}
  > cvg3[1]
  $ERR127306
  $ERR127306[[1]]
  integer-Rle of length 21469908 with 489540 runs
    Lengths: 6818    9    8    1    1    2    2 ...    3    5    8    1   10  863
    Values :    0   22   23   19   17   18   17 ...   20   22   21   23   22    0
\end{verbatim}

\newpage
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
\section{Chunking}
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%

\subsection{Ranges in a file}

Both \Rcode{reduceByFile} and \Rcode{reduceByRange} process \Rcode{ranges} one
element at a time. When \Rcode{ranges} is a GRanges the element is a single
range and when it is a GRangesList the element can contain multiple ranges.

If the GRanges is very long (many ranges) working one range at a time can be
inefficient. Splitting the GRanges into a GRangesList allows
\Rcode{reduceByFile} and \Rcode{reduceByRange} to work on groups of ranges and
will gain speed and efficiency in most applications. This approach works as long
as the analysis does not depend on keeping the ranges separate (i.e., MAP and
REDUCE can be written to operate on groups of ranges instead of a single range).

For applications that combine data \emph{within} a file, chunking can be done
with \Rcode{reduceByFile} and a GRangesList.  Similarly, when chunking through
ranges to combine data \emph{across} files use \Rcode{reduceByRange} with a
GRangesList.


\subsection{Records in a file}

\Rcode{reduceByYield} iterates through records in a single file
that would otherwise not fit in memory. It is similar to a one dimensional 
\Rcode{reduceByFile} but the arguments and approach are slightly different.

Similar to other \Rcode{GenomicFiles} functions, data are manipulated and 
reduced with \Rcode{MAP} and \Rcode{REDUCE} functions. What sets
\Rcode{reduceByYield} apart are the use of \Rcode{YIELD} and \Rcode{DONE}
arguments. \Rcode{YIELD} is a function that returns a chunk of data
to work on and \Rcode{DONE} is a function that defines a stopping criteria.

Records from a single file are read by \Rcode{readGAlignments} and limited by 
the \Rcode{yieldSize} set in the BamFile. 
<<reduceByYield-YIELD>>=
library(GenomicAlignments)
bf <- BamFile(fls[1], yieldSize=100000)
YIELD <- function(x, ...) readGAlignments(x)
@

MAP counts overlaps between the reads and a GRanges of interest while REDUCE 
sums counts over the chunks.
<<reduceByYield-MAP-REDUCE>>=
gr <- unlist(tiles, use.names=FALSE)
MAP <- function(value, gr, ...) {
    requireNamespace("GenomicRanges")  ## for countOverlaps()
    GenomicRanges::countOverlaps(gr, value)
}
REDUCE <- `+` 
@

When \Rcode{DONE} evaluates to TRUE, iteration stops. `value` is the
object returned from calling YIELD on the BAM file. At the end of file
the length of records will be 0 and \Rcode{DONE} will evaluate to TRUE.
<<reduceByYield-DONE>>=
DONE <- function(value) length(value) == 0L
@

The MAP step is run in parallel when \Rcode{parallel=TRUE}. `parallel` is
currently implemented for Unix/Mac only so we use multicore workers.
\begin{verbatim}
register(MulticoreParam(3))
> reduceByYield(bf, YIELD, MAP, REDUCE, DONE, gr=gr, parallel=TRUE)
[[1]]
[1]  21465 163154  75498 212593 327785
\end{verbatim}

Taking this one step further, we can use \Rcode{bplapply} to distribute files to
workers and call \Rcode{reduceByYield} on each file. If adequate resources are
available this example could have 2 levels of parallel dispatch, one at the file
level (\Rcode{bplapply}) and one at the MAP level (\Rcode{reduceByYield(...,
parallel=TRUE)}. This example takes the conservative approach and runs
\Rcode{reduceByYield} in serial on each worker.

The function `FUN` will be run on each worker.
<<reduceByYield-bplapply>>=
FUN <- function(file, gr, YIELD, MAP, REDUCE, tiles, ...) {
    requireNamespace("GenomicAlignments")  ## for BamFile, readGAlignments()
    requireNamespace("GenomicFiles")       ## for reduceByYield()
    gr <- unlist(tiles, use.names=FALSE)
    bf <- Rsamtools::BamFile(file, yieldSize=100000)
    YIELD <- function(x, ...) GenomicAlignments::readGAlignments(x)
    MAP <- function(value, gr, ...) {
        requireNamespace("GenomicRanges")  ## for countOverlaps()
        GenomicRanges::countOverlaps(gr, value)
    }
    REDUCE <- `+` 
    GenomicFiles::reduceByYield(bf, YIELD, MAP, REDUCE, gr=gr, parallel=FALSE)
}
@

\Rcode{bplapply} distributes the files to workers. Each worker uses
\Rcode{reduceByYield} to iteratively count and reduce overlaps in a BAM file.

\begin{verbatim}
  > bplapply(fls, FUN, gr=gr, YIELD=YIELD, MAP=MAP, REDUCE=REDUCE, tiles=tiles)
  $ERR127306
  [1]  21465 163154  75498 212593 327785

  $ERR127307
  [1]  23544 181551  91702 236845 341670
 
  $ERR127308
  [1]  23236 178270  84027 234735 355353
 
  $ERR127309
  [1]  20890 160804  82120 208961 305701
 
  $ERR127302
  [1]  20636 140052  89834 208824 283432
 
  $ERR127303
  [1]  22198 149809 106987 226217 281000
 
  $ERR127304
  [1]  25718 150984  94198 223797 316043
 
  $ERR127305
  [1]  25646 145655  79854 219333 327909
\end{verbatim}


%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
\section{\Rcode{sessionInfo()}}
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%

<<sessionInfo, results=tex>>=
toLatex(sessionInfo())
@

\end{document}
