## ----setup, include = FALSE---------------------------------------------------
library(RiboDiPA)
knitr::opts_chunk$set(
    collapse = TRUE,
    comment = "#>"
)

## ---- warning=FALSE, message=FALSE, eval=FALSE--------------------------------
#  if(!requireNamespace("BiocManager", quietly = TRUE))
#      install.packages("BiocManager")
#  BiocManager::install("RiboDiPA")

## ---- warning=FALSE, message=FALSE--------------------------------------------
## Download sample files from GitHub
library(BiocFileCache)
file_names <- c("WT1.bam", "WT2.bam", "MUT1.bam", "MUT2.bam", "eg.gtf")
url <- "https://github.com/jipingw/RiboDiPA-data/raw/master/"
bfc <- BiocFileCache()
bam_path <- bfcrpath(bfc,paste0(url,file_names))

## ---- warning=FALSE, message=FALSE--------------------------------------------
## Make the class label for the experiment
classlabel <- data.frame(
    condition = c("mutant", "mutant", "wildtype", "wildtype"),
    comparison = c(2, 2, 1, 1)
)
rownames(classlabel) <- c("mutant1","mutant2","wildtype1","wildtype2")

## ---- warning=FALSE, message=FALSE--------------------------------------------
## Run the RiboDiPA pipeline with default parameters
result.wrp <- RiboDiPA(bam_path[1:4], bam_path[5], classlabel, cores = 2)

## -----------------------------------------------------------------------------
## View the table of output from RiboDiPA
head(result.wrp$gene[order(result.wrp$gene$qvalue), ])

## ---- warning=FALSE, message=FALSE--------------------------------------------
## Perform individual P-site mapping procedure
data.psite <- psiteMapping(bam_file_list = bam_path[1:4], 
    gtf_file = bam_path[5], psite.mapping = "auto", cores = 2)

## -----------------------------------------------------------------------------
## P-site mapping offset rule generated
data.psite$psite.mapping

## ---- warning=FALSE, message=FALSE, eval=FALSE--------------------------------
#  ## Use user specified psite mapping offset rule
#  offsets <- cbind(qwidth = c(28, 29, 30, 31, 32),
#      psite = c(18, 18, 18, 19, 19))
#  data.psite2 <- psiteMapping(bam_path[1:4], bam_path[5],
#      psite.mapping = offsets, cores = 2)

## ---- warning=FALSE, message=FALSE,eval=FALSE---------------------------------
#  ## Merge the P-site data into bins with a fixed or an adaptive width
#  data.binned <- dataBinning(data = data.psite$coverage, bin.width = 0,
#      zero.omit = FALSE, bin.from.5UTR = TRUE, cores = 2)

## ---- warning=FALSE, message=FALSE,eval=FALSE---------------------------------
#  ## Merge the P-site data on each codon
#  data.codon <- dataBinning(data = data.psite$coverage, bin.width = 1,
#      zero.omit = FALSE, bin.from.5UTR = TRUE, cores = 2)

## ---- warning=FALSE, message=FALSE,eval=FALSE---------------------------------
#  ## Merge the P-site data on each exon and perform differential pattern analysis
#  result.exon <- diffPatternTestExon(psitemap = data.psite,
#      classlabel = classlabel, method = c('gtxr', 'qvalue'))

## ---- warning=FALSE, message=FALSE,eval=FALSE---------------------------------
#  ## Perform differential pattern analysis
#  result.pst <- diffPatternTest(data = data.binned,
#      classlabel = classlabel, method=c('gtxr', 'qvalue'))

## ----fig2, fig.height=6, fig.width=6, fig.align="center", results='hide'------
## Plot ribosome per nucleotide tracks of specified genes.
plotTrack(data = data.psite, genes.list = c("YDR050C", "YDR064W"),
    replicates = NULL, exons = FALSE)

## ----fig3, fig.height = 9, fig.width = 10, fig.align = "center",results='hide'----
## Plot binned ribosome tracks of siginificant genes: YDR086C and YDR210W.
## you can specify the thrshold to redefine the significant level
plotTest(result = result.pst, genes.list = NULL, threshold = 0.05) 

## ---- warning=FALSE, message=FALSE, eval=FALSE--------------------------------
#  ##base-bair RPF track
#  library(igvR)
#  thred <- 0.05
#  igv <- igvR()
#  setBrowserWindowTitle(igv, "ribosome footprint track example")
#  setGenome(igv, "saccer3")
#  
#  data(data.psite)
#  names.rep <- c("mutant1", "mutant2", "wildtype1", "wildtype2")
#  tracks.bp <- bpTrack(data = data.psite, names.rep = names.rep,
#      genes.list = c("YDR050C", "YDR062W", "YDR064W"))
#  
#  for(track.name in names.rep){
#      track.rep <- tracks.bp[[track.name]]
#      track <- GRangesQuantitativeTrack(trackName = paste(track.name, "bp"),
#          track.rep[,1], color = "green")
#      displayTrack(igv, track)
#  }}
#  

## ---- warning=FALSE, message=FALSE, eval=FALSE--------------------------------
#  ## bin track and test results
#  data(result.pst)
#  data(data.psite)
#  tracks.bin <- binTrack(data = result.pst, exon.anno = data.psite$exons)
#  
#  for(track.name in names(tracks.bin)){
#      track.rep <- tracks.bin[[track.name]]
#      resize(track.rep, width(track.rep) + 1)
#      track <- GRangesQuantitativeTrack(trackName = paste(track.name, "binned"),
#          track.rep[,1], color = "black")
#      displayTrack(igv, track)
#  }
#  
#  track.rep2 <- tracks.bin[[1]]
#  sig.bin <- (values(track.rep2)[,5] <= thred)
#  log10.padj <- - log10(values(track.rep2)[,5])
#  mcols(track.rep2) <- data.frame(log10padj = log10.padj)
#  track.rep2 <- track.rep2[which(sig.bin),]
#  track <- GRangesQuantitativeTrack(trackName = "- log 10 of padj",
#      track.rep2, color = "red", trackHeight = 40)
#  displayTrack(igv, track)

## ---- warning=FALSE, message=FALSE, eval=FALSE--------------------------------
#  ## bin track and test results
#  igv2 <- igvR()
#  setBrowserWindowTitle(igv2, "ribosome footprint per exon example")
#  setGenome(igv2, "saccer3")
#  data(result.exon)
#  tracks.exon <- exonTrack(data = result.exon, gene = "tY(GUA)D")
#  for(track.name in names(tracks.exon)){
#      track.rep <- tracks.exon[[track.name]]
#      for(tx.name in names(track.rep)){
#          track.tx <- tracks.exon[[track.name]][[tx.name]]
#          track <- GRangesQuantitativeTrack(trackName =
#              paste(track.name, tx.name), track.tx[,1], color = track.name)
#          displayTrack(igv2, track)
#      }
#  }

## ----sessionInfo--------------------------------------------------------------
sessionInfo()

