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#' Convert phyloseq data to DESeq2 dds object
#'
#' No testing is performed by this function. The phyloseq data is converted
#' to the relevant \code{\link[DESeq2]{DESeqDataSet}} object, which can then be
#' tested in the negative binomial generalized linear model framework
#' of the \code{\link[DESeq2]{DESeq}} function in DESeq2 package.
#' See the
#' \href{http://joey711.github.io/phyloseq-extensions}{phyloseq-extensions}
#' tutorials for more details.
#'
#' @param physeq (Required). \code{\link{phyloseq-class}}.
#' Must have a \code{\link{sample_data}} component.
#'
#' @param design (Required). A \code{\link{formula}} which specifies the design of the experiment,
#' taking the form \code{formula(~ x + y + z)}. That is, a formula with right-hand side only.
#' By default, the functions in this package and DESeq2
#' will use the last variable in the formula (e.g. \code{z})
#' for presenting results (fold changes, etc.) and plotting.
#' When considering your specification of experimental design, you will want to
#' re-order the levels so that the \code{NULL} set is first.
#' For example, the following line of code would ensure that Enterotype 1 is used as the
#' reference sample class in tests by setting it to the first of the factor levels
#' using the \code{\link{relevel}} function:
#'
#' \code{sample_data(entill)$Enterotype <- relevel(sample_data(entill)$Enterotype, "1")}
#'
#' @param ... (Optional). Additional named arguments passed to \code{\link[DESeq2]{DESeqDataSetFromMatrix}}.
#' Most users will not need to pass any additional arguments here.
#' Most testing-related options should be provided in
#' a following call to \code{\link[DESeq2]{DESeq}}.
#'
#' @return A \code{\link[DESeq2]{DESeqDataSet}} object.
#'
#' @seealso
#'
#' \code{vignette("phyloseq-mixture-models")}
#'
#' The
#' \href{http://joey711.github.io/phyloseq-extensions}{phyloseq-extensions}
#' tutorials.
#'
#' \code{\link[DESeq2]{DESeq}}
#'
#' \code{\link[DESeq2]{results}}
#'
#' \code{\link[DESeq2]{DESeqDataSetFromMatrix}}
#'
#' @export
#'
#' @examples
#' # Check out the vignette phyloseq-mixture-models for more details.
#' # vignette("phyloseq-mixture-models")
#' data(soilrep)
#' phyloseq_to_deseq2(soilrep, ~warmed)
phyloseq_to_deseq2 = function(physeq, design, ...){
# Need to add check here for missing sample_data
if( is.null(sample_data(physeq, FALSE)) ){
stop("There must be sample_data present, for specifying experimental design. See ?phyloseq_to_deseq2")
}
# Enforce orientation. Samples are columns
if( !taxa_are_rows(physeq) ){ physeq <- t(physeq)}
# Coerce count data to vanilla matrix of integers
countData = round(as(otu_table(physeq), "matrix"), digits=0)
colData = data.frame(sample_data(physeq))
# Create the DESeq data set, dds.
if(requireNamespace("DESeq2")){
dds <- DESeq2::DESeqDataSetFromMatrix(countData, colData, design, ...)
return(dds)
}
}
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