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Commit made by the Bioconductor Git-SVN bridge.

Commit id: 21404fe8275c89f6f29231a680703827b0796ff9

Merge pull request #4 from PeteHaitch/master

Updates to read.bismark(), mostly documentation

Commit id: 6ba560784b0e6e6f2e8c40de72b4abf516dc14e3

Update tests and examples to use gzipped data. Ensure strand is kept for cytosineReport files if collapseStrand = FALSE.


Commit id: 41b003506ba63d0c7ec330c2ea0b826e0c1c5e9c

Remove Travis-CI. Not relevant at this juncture.


Commit id: 2fd84392d9f39409b6958f0ce0d77ea5e6476bee

Hardcode RVERSION in .travis.yml


Commit id: 127bc93e92cd065e94dbeac0001949e5c4138d91

Experimental Travis-CI support.


Commit id: a6fecb9692f9b72093d3e0e79cf39e325d63b1e9

Second pass at updating read.bismark() and docs. Updated DESCRIPTION in light of changes and added NEWS.rd entry.


Commit id: 5af9b047653567816174480a6f405e1d02f0a74b

First pass at re-write of read.bismark() and associated docs.



git-svn-id: https://hedgehog.fhcrc.org/bioconductor/trunk/madman/Rpacks/bsseq@106909 bc3139a8-67e5-0310-9ffc-ced21a209358

khansen authored on 30/07/2015 01:16:09
Showing 13 changed files

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new file mode 100644
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+^.*\.Rproj$
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+^\.Rproj\.user$
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new file mode 100644
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+.Rproj.user
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+.Rhistory
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+.RData
... ...
@@ -1,17 +1,52 @@
1 1
 Package: bsseq
2
-Version: 1.5.4
2
+Version: 1.5.5
3 3
 Title: Analyze, manage and store bisulfite sequencing data
4
-Description: A collection of tools for analyzing and visualizing bisulfite sequencing data.
4
+Description: A collection of tools for analyzing and visualizing bisulfite
5
+    sequencing data.
5 6
 Authors@R: c(person(c("Kasper", "Daniel"), "Hansen", role = c("aut", "cre"),
6
-                    email = "kasperdanielhansen@gmail.com"))
7
-Depends: R (>= 2.15), methods, BiocGenerics, matrixStats, IRanges (>= 2.1.10),
8
-        GenomicRanges (>= 1.19.6), SummarizedExperiment (>= 0.1.1), parallel,
9
-        GenomeInfoDb
10
-Imports: scales, stats, graphics, Biobase, locfit, gtools, data.table, S4Vectors
11
-Suggests: RUnit, bsseqData, BiocStyle
12
-Collate: hasGRanges.R BSseq_class.R BSseqTstat_class.R BSseq_utils.R combine.R 
13
-        utils.R read.bsmooth.R read.bismark.R BSmooth.R BSmooth.tstat.R dmrFinder.R
14
-        gof_stats.R plotting.R fisher.R permutations.R
7
+    email = "kasperdanielhansen@gmail.com"),
8
+    person("Peter", "Hickey", role = "ctb", email = "peter.hickey@gmail.com"))
9
+Depends:
10
+    R (>= 2.15),
11
+    methods,
12
+    BiocGenerics,
13
+    IRanges (>= 2.1.10),
14
+    GenomicRanges (>= 1.19.6),
15
+    SummarizedExperiment (>= 0.1.1),
16
+    parallel,
17
+    GenomeInfoDb
18
+Imports:
19
+    scales,
20
+    stats,
21
+    graphics,
22
+    Biobase,
23
+    locfit,
24
+    gtools,
25
+    data.table,
26
+    S4Vectors,
27
+    R.utils (>= 2.0.0),
28
+    matrixStats
29
+Suggests:
30
+    RUnit,
31
+    bsseqData,
32
+    BiocStyle
33
+Collate:
34
+    hasGRanges.R
35
+    BSseq_class.R
36
+    BSseqTstat_class.R
37
+    BSseq_utils.R
38
+    combine.R
39
+    utils.R
40
+    read.bsmooth.R
41
+    read.bismark.R
42
+    BSmooth.R
43
+    BSmooth.tstat.R
44
+    dmrFinder.R
45
+    gof_stats.R
46
+    plotting.R
47
+    fisher.R
48
+    permutations.R
49
+    BSmooth.fstat.R
15 50
 License: Artistic-2.0
16 51
 URL: https://github.com/kasperdanielhansen/bsseq
17 52
 LazyData: yes
... ...
@@ -30,6 +30,7 @@ importMethodsFrom(GenomeInfoDb, "seqlengths", "seqlengths<-", "seqinfo", "seqinf
30 30
 import(S4Vectors)
31 31
 importFrom(gtools, "combinations")
32 32
 importFrom(data.table, "fread", "setnames")
33
+importFrom(R.utils, "isGzipped", "gunzip")
33 34
 
34 35
 ##
35 36
 ## Exporting
... ...
@@ -60,7 +61,7 @@ export("BSseq", "getMeth", "getCoverage", "getBSseq", "getStats",
60 61
        "collapseBSseq", "orderBSseq", "hasBeenSmoothed", "chrSelectBSseq",
61 62
        "BSmooth", "BSmooth.tstat", "dmrFinder", "fisherTests",
62 63
        "combineList", "strandCollapse",
63
-       "plotRegion", "plotManyRegions", 
64
+       "plotRegion", "plotManyRegions",
64 65
        "read.umtab", "read.umtab2", "read.bsmooth", "read.bismark",
65 66
        "poissonGoodnessOfFit", "binomialGoodnessOfFit",
66 67
        "data.frame2GRanges", "BSseqTstat")
67 68
new file mode 100644
... ...
@@ -0,0 +1,138 @@
1
+BSmooth.fstat <- function(BSseq, group1, group2, estimate.var = c("same", "paired", "group2"),
2
+                          local.correct = TRUE, maxGap = NULL, qSd = 0.75, k = 101, mc.cores = 1, verbose = TRUE){
3
+    smoothSd <- function(Sds, k) {
4
+        k0 <- floor(k/2)
5
+        if(all(is.na(Sds))) return(Sds)
6
+        thresSD <- pmax(Sds, quantile(Sds, qSd, na.rm = TRUE), na.rm = TRUE)
7
+        addSD <- rep(median(Sds, na.rm = TRUE), k0)
8
+        sSds <- as.vector(runmean(Rle(c(addSD, thresSD, addSD)), k = k))
9
+        sSds
10
+    }
11
+    compute.correction <- function(idx, qSd = 0.75) {
12
+        xx <- start(BSseq)[idx]
13
+        yy <- tstat[idx]
14
+        suppressWarnings({
15
+            drange <- diff(range(xx, na.rm = TRUE))
16
+        })
17
+        if(drange <= 25000)
18
+            return(yy)
19
+        tstat.function <- approxfun(xx, yy)
20
+        xx.reg <- seq(from = min(xx), to = max(xx), by = 2000)
21
+        yy.reg <- tstat.function(xx.reg)
22
+        fit <- locfit(yy.reg ~ lp(xx.reg, h = 25000, deg = 2, nn = 0),
23
+                      family = "huber", maxk = 50000) 
24
+        correction <- predict(fit, newdata = data.frame(xx.reg = xx))
25
+        yy - correction 
26
+    }
27
+
28
+    estimate.var <- match.arg(estimate.var)
29
+    stopifnot(is(BSseq, "BSseq"))
30
+    stopifnot(hasBeenSmoothed(BSseq))
31
+    if(is.character(group1)) {
32
+        stopifnot(all(group1 %in% sampleNames(BSseq)))
33
+        group1 <- match(group1, sampleNames(BSseq))
34
+    }
35
+    if(is.numeric(group1)) {
36
+        stopifnot(min(group1) >= 1 & max(group1) <= ncol(BSseq))
37
+    } else stop("problems with argument 'group1'")
38
+    if(is.character(group2)) {
39
+        stopifnot(all(group2 %in% sampleNames(BSseq)))
40
+        group2 <- match(group2, sampleNames(BSseq))
41
+    }    
42
+    if(is.numeric(group2)) {
43
+        stopifnot(min(group2) >= 1 & max(group2) <= ncol(BSseq))
44
+    } else stop("problems with argument 'group2'")
45
+    stopifnot(length(intersect(group1, group2)) == 0)
46
+    stopifnot(length(group1) > 0)
47
+    stopifnot(length(group2) > 0)
48
+    stopifnot(length(group1) + length(group2) >= 3)
49
+    if(estimate.var == "paired")
50
+        stopifnot(length(group1) == length(group2))
51
+    
52
+    if(any(rowSums(getCoverage(BSseq)[, c(group1, group2)]) == 0))
53
+        warning("Computing f-statistics at locations where there is no data; consider subsetting the 'BSseq' object first")
54
+    
55
+    if(is.null(maxGap))
56
+        maxGap <- BSseq@parameters$maxGap
57
+    if(is.null(maxGap))
58
+        stop("need to set argument 'maxGap'")
59
+    
60
+    if(verbose) cat("[BSmooth.fstat] preprocessing ... ")
61
+    ptime1 <- proc.time()
62
+    clusterIdx <- makeClusters(BSseq, maxGap = maxGap)
63
+    ptime2 <- proc.time()
64
+    stime <- (ptime2 - ptime1)[3]
65
+    if(verbose) cat(sprintf("done in %.1f sec\n", stime))
66
+        
67
+    if(verbose) cat("[BSmooth.fstat] computing stats within groups ... ")
68
+    ptime1 <- proc.time()
69
+    allPs <- getMeth(BSseq, type = "smooth", what = "perBase",
70
+                     confint = FALSE)
71
+    group1.means <- rowMeans(allPs[, group1, drop = FALSE], na.rm = TRUE)
72
+    group2.means <- rowMeans(allPs[, group2, drop = FALSE], na.rm = TRUE)
73
+    ptime2 <- proc.time()
74
+    stime <- (ptime2 - ptime1)[3]
75
+    if(verbose) cat(sprintf("done in %.1f sec\n", stime))
76
+    
77
+    if(verbose) cat("[BSmooth.fstat] computing stats across groups ... ")
78
+    ptime1 <- proc.time()
79
+    switch(estimate.var,
80
+           "group2" = {
81
+               rawSds <- rowSds(allPs[, group2, drop = FALSE], na.rm = TRUE)
82
+               smoothSds <- do.call(c, mclapply(clusterIdx, function(idx) {
83
+                   smoothSd(rawSds[idx], k = k)
84
+               }, mc.cores = mc.cores))
85
+               scale <- sqrt(1/length(group1) + 1/length(group2))
86
+               tstat.sd <- smoothSds * scale
87
+           },
88
+           "same" = {
89
+               rawSds <- sqrt( ((length(group1) - 1) * rowVars(allPs[, group1, drop = FALSE]) +
90
+                                (length(group2) - 1) * rowVars(allPs[, group2, drop = FALSE])) /
91
+                              (length(group1) + length(group2) - 2))
92
+               smoothSds <- do.call(c, mclapply(clusterIdx, function(idx) {
93
+                   smoothSd(rawSds[idx], k = k)
94
+               }, mc.cores = mc.cores))
95
+               scale <- sqrt(1/length(group1) + 1/length(group2))
96
+               tstat.sd <- smoothSds * scale
97
+           },
98
+           "paired" = {
99
+               rawSds <- rowSds(allPs[, group1, drop = FALSE] - allPs[, group2, drop = FALSE])
100
+               smoothSds <- do.call(c, mclapply(clusterIdx, function(idx) {
101
+                   smoothSd(rawSds[idx], k = k)
102
+               }, mc.cores = mc.cores))
103
+               scale <- sqrt(1/length(group1))
104
+               tstat.sd <- smoothSds * scale
105
+           })
106
+    tstat <- (group1.means - group2.means) / tstat.sd
107
+    is.na(tstat)[tstat.sd == 0] <- TRUE
108
+    if(local.correct) {
109
+        tstat.corrected <- do.call(c, mclapply(clusterIdx,
110
+                                               compute.correction, qSd = qSd,
111
+                                               mc.cores = mc.cores))
112
+    }
113
+    ptime2 <- proc.time()
114
+    stime <- (ptime2 - ptime1)[3]
115
+    if(verbose) cat(sprintf("done in %.1f sec\n", stime))
116
+    
117
+    if(local.correct) {
118
+        stats <- cbind(rawSds, tstat.sd, group2.means, group1.means,
119
+                       tstat, tstat.corrected)
120
+        colnames(stats) <- c("rawSds", "tstat.sd",
121
+                             "group2.means", "group1.means", "tstat",
122
+                             "tstat.corrected")
123
+ 
124
+    } else {
125
+        stats <- cbind(rawSds, tstat.sd, group2.means, group1.means,
126
+                       tstat)
127
+        colnames(stats) <- c("rawSds", "tstat.sd",
128
+                             "group2.means", "group1.means", "tstat")
129
+    }
130
+    
131
+    parameters <- c(BSseq@parameters,
132
+                    list(tstatText = sprintf("BSmooth.fstat (local.correct = %s, maxGap = %d)",
133
+                         local.correct, maxGap),
134
+                         group1 = group1, group2 = group2, k = k, qSd = qSd,
135
+                         local.correct = local.correct, maxGap = maxGap))
136
+    out <- BSseqTstat(gr = granges(BSseq), stats = stats, parameters = parameters)
137
+    out
138
+}
... ...
@@ -1,98 +1,125 @@
1
-read.bismark <- function(files, sampleNames, rmZeroCov = FALSE, strandCollapse = TRUE,
2
-                         fileType = c("cytosineReport", "oldBedGraph"), verbose = TRUE){
1
+read.bismark <- function(files,
2
+                         sampleNames,
3
+                         rmZeroCov = FALSE,
4
+                         strandCollapse = TRUE,
5
+                         fileType = c("cov", "oldBedGraph", "cytosineReport"),
6
+                         verbose = TRUE) {
3 7
     ## Argument checking
4
-    if (anyDuplicated(files)){
8
+    if (anyDuplicated(files)) {
5 9
         stop("duplicate entries in 'files'")
6 10
     }
7
-    if (length(sampleNames) != length(files) | anyDuplicated(sampleNames)){
11
+    if (length(sampleNames) != length(files) || anyDuplicated(sampleNames)) {
8 12
         stop("argument 'sampleNames' has to have the same length as argument 'files', without duplicate entries")
9 13
     }
10 14
     fileType <- match.arg(fileType)
15
+    if (verbose) {
16
+        message(paste0("Assuming file type is", fileType))
17
+    }
11 18
     ## Process each file
12 19
     idxes <- seq_along(files)
13 20
     names(idxes) <- sampleNames
14
-    allOut <- lapply(idxes, function(ii){
21
+    allOut <- lapply(idxes, function(ii) {
15 22
         if (verbose) {
16 23
             cat(sprintf("[read.bismark] Reading file '%s' ... ", files[ii]))
17 24
         }
18 25
         ptime1 <- proc.time()
19
-        if(fileType == "oldBedGraph") {
20
-            raw <- read.bismarkFileRaw(thisfile = files[ii])
21
-        }
22
-        if(fileType == "cytosineReport") {
23
-            raw <- read.bismarkCytosineRaw(thisfile = files[ii], keepContext = FALSE)
26
+        if (fileType == "cov" || fileType == "oldBedGraph") {
27
+            out <- read.bismarkCovRaw(thisfile = files[ii],
28
+                                      thisSampleName = sampleNames[ii],
29
+                                      rmZeroCov = rmZeroCov)
30
+        } else if (fileType == "cytosineReport") {
31
+            out <- read.bismarkCytosineReportRaw(thisfile = files[ii],
32
+                                                 thisSampleName = sampleNames[ii],
33
+                                                 rmZeroCov = rmZeroCov,
34
+                                                 keepContext = FALSE)
24 35
         }
25
-        M <- matrix(mcols(raw)[, "mCount"], ncol = 1)
26
-        Cov <- M + mcols(raw)[, "uCount"]
27
-        mcols(raw) <- NULL
28
-        out <- BSseq(gr = raw, M = M, Cov = Cov,
29
-                     sampleNames = sampleNames[ii], rmZeroCov = rmZeroCov)
30
-        if(strandCollapse && !all(runValue(strand(out)) == "*"))
36
+        if (strandCollapse && !all(runValue(strand(out)) == "*")) {
31 37
             out <- strandCollapse(out)
38
+        }
32 39
         ptime2 <- proc.time()
33 40
         stime <- (ptime2 - ptime1)[3]
34 41
         if (verbose) {
35
-            cat(sprintf("done in %.1f secs\n", stime))  
42
+            cat(sprintf("done in %.1f secs\n", stime))
36 43
         }
37
-        out  
44
+        out
38 45
     })
46
+
39 47
     if (verbose) {
40 48
         cat(sprintf("[read.bismark] Joining samples ... "))
41 49
     }
42 50
     ptime1 <- proc.time()
43 51
     allOut <- combineList(allOut)
44 52
     ptime2 <- proc.time()
45
-    stime <- (ptime2 - ptime1)[3]
53
+    stime <- (ptime2 - ptime1)[3L]
46 54
     if (verbose) {
47 55
         cat(sprintf("done in %.1f secs\n", stime))
48 56
     }
49 57
     allOut
50 58
 }
51 59
 
52
-read.bismarkFileRaw <- function(thisfile, verbose = TRUE){
53
-    ## Set up the 'what' argument for scan()
54
-    columnHeaders <- c("chr", "start", "end", "mPerc", "mCount", "uCount")
55
-    what0 <- replicate(length(columnHeaders), character(0))
56
-    names(what0) <- columnHeaders
57
-    int <- which(columnHeaders %in% c("start", "end", "mCount", "uCount"))
58
-    what0[int] <- replicate(length(int), integer(0))
59
-    null <- which(columnHeaders %in% "mPerc")
60
-    what0[null] <- replicate(length(null), NULL)
60
+read.bismarkCovRaw <- function(thisfile,
61
+                               thisSampleName,
62
+                               rmZeroCov) {
63
+
64
+    ## data.table::fread() can't read directly from a gzipped file so, if
65
+    ## necessary, gunzip the file to a temporary location.
66
+    if (isGzipped(thisfile)) {
67
+        thisfile <- gunzip(thisfile,
68
+                           temporary = TRUE,
69
+                           overwrite = TRUE,
70
+                           remove = FALSE)
71
+    }
72
+
61 73
     ## Read in the file
62
-    if (grepl("\\.gz$", thisfile)) 
63
-        con <- gzfile(thisfile)
64
-    else 
65
-        con <- file(thisfile, open = "r")
66
-    out <- scan(file = con, what = what0, sep = "\t", quote = "", na.strings = "NA", quiet = TRUE)
67
-    close(con)
74
+    out <- fread(thisfile)
75
+    if (ncol(out) != 6L) {
76
+        stop("unknown file format")
77
+    }
78
+
68 79
     ## Create GRanges instance from 'out'
69
-    gr <- GRanges(seqnames = out[["chr"]], ranges = IRanges(start = out[["start"]], width = 1))
70
-    out[["chr"]] <- out[["start"]] <- out[["end"]] <- NULL
71
-    out <- out[!sapply(out, is.null)]
72
-    df <- DataFrame(out)
73
-    mcols(gr) <- df
74
-    gr
80
+    gr <- GRanges(seqnames = out[[1L]],
81
+                  ranges = IRanges(start = out[[2L]], width = 1L))
82
+
83
+    ## Create BSseq instance from 'out'
84
+    BSseq(gr = gr,
85
+          M = as.matrix(out[[5L]]),
86
+          Cov = as.matrix(out[[5L]] + out[[6L]]),
87
+          sampleNames = thisSampleName,
88
+          rmZeroCov = rmZeroCov)
75 89
 }
76 90
 
77
-read.bismarkCytosineRaw <- function(thisfile, keepContext = FALSE) {
91
+read.bismarkCytosineReportRaw <- function(thisfile,
92
+                                          thisSampleName,
93
+                                          rmZeroCov,
94
+                                          keepContext = FALSE) {
95
+
96
+    ## NOTE: keepContext not yet implemented
97
+    if (keepContext) {
98
+        stop("'keepContext' argument not yet implemented")
99
+    }
100
+
101
+    ## data.table::fread() can't read directly from a gzipped file so, if
102
+    ## necessary, gunzip the file to a temporary location.
103
+    if (isGzipped(thisfile)) {
104
+        thisfile <- gunzip(thisfile,
105
+                           temporary = TRUE,
106
+                           overwrite = TRUE,
107
+                           remove = FALSE)
108
+    }
109
+
110
+    ## Read in the file
78 111
     out <- fread(thisfile)
79
-    if(length(out) != 6 && length(out) != 7)
112
+    if (ncol(out) != 7L) {
80 113
         stop("unknown file format")
81
-    if(length(out) == 6) {
82
-        setnames(out, c("chr", "start", "end", "methPerc", "mCount", "uCount"))
83
-        gr <- GRanges(seqnames = out[["chr"]],
84
-                      ranges = IRanges(start = out[["start"]], width = 1),
85
-                      mCount = out[["mCount"]], uCount = out[["uCount"]])
86
-    }
87
-    if(length(out) == 7) {
88
-        setnames(out, c("chr", "start", "strand", "mCount", "uCount", "type", "context"))
89
-        gr <- GRanges(seqnames = out[["chr"]], strand = out[["strand"]],
90
-                      ranges = IRanges(start = out[["start"]], width = 1),
91
-                      mCount = out[["mCount"]], uCount = out[["uCount"]])
92
-        if(keepContext) {
93
-            gr$type <- out[["type"]]
94
-            gr$context <- out[["context"]]
95
-        }                      
96 114
     }
97
-    gr
115
+
116
+    ## Create GRanges instance from 'out'
117
+    gr <- GRanges(seqnames = out[[1]],
118
+                  ranges = IRanges(start = out[[2]], width = 1),
119
+                  strand = out[[3]])
120
+
121
+    ## Create BSseq instance from 'out'
122
+    BSseq(gr = gr,
123
+          M = as.matrix(out[[4L]]),
124
+          Cov = as.matrix(out[[4L]] + out[[5L]]))
98 125
 }
99 126
new file mode 100644
... ...
@@ -0,0 +1,20 @@
1
+Version: 1.0
2
+
3
+RestoreWorkspace: Default
4
+SaveWorkspace: Default
5
+AlwaysSaveHistory: Default
6
+
7
+EnableCodeIndexing: Yes
8
+UseSpacesForTab: Yes
9
+NumSpacesForTab: 4
10
+Encoding: UTF-8
11
+
12
+RnwWeave: knitr
13
+LaTeX: pdfLaTeX
14
+
15
+AutoAppendNewline: Yes
16
+StripTrailingWhitespace: Yes
17
+
18
+BuildType: Package
19
+PackageUseDevtools: Yes
20
+PackageInstallArgs: --no-multiarch --with-keep.source
... ...
@@ -6,8 +6,11 @@
6 6
   \itemize{
7 7
     \item new function strandCollapse for collapsing forward and reverse
8 8
     strand data to be unstranded.
9
-    \item Updated read.bismark to support the cytosine report files;
10
-    both formats are support.
9
+    \item Updated read.bismark() to support the cytosine report files;
10
+    both formats are supported. Other minor updates (mostly internal) to
11
+    read.bismark(). Greatly improved documentation of this function, paying
12
+    particular attention to differences in file formats between versions of
13
+    Bismark.
11 14
   }
12 15
 }
13 16
 
... ...
@@ -103,7 +106,7 @@
103 106
     combineList for testing.
104 107
     \item Bug fix to read.bsmooth; it now works correctly for the
105 108
     default settings (= returning a single object of class BSseq and not a
106
-    list). 
109
+    list).
107 110
     \item Getting ready for initial release on Bioconductor.
108 111
     \item Updated the citations in the vignette(s) and the CITATION file.
109 112
   }
110 113
deleted file mode 100644
... ...
@@ -1,2013 +0,0 @@
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