extractPatterns: extractPatterns

In BBCG/epialleleR: Fast, Epiallele-Aware Methylation Caller and Reporter

extractPatterns

Description

This function extracts methylation patterns (epialleles) for a given genomic region of interest.

Usage

extractPatterns(
  bam,
  bed,
  bed.row = 1,
  zero.based.bed = FALSE,
  match.min.overlap = 1,
  extract.context = c("CG", "CHG", "CHH", "CxG", "CX"),
  min.context.freq = 0.01,
  clip.patterns = FALSE,
  strand.offset = c(CG = 1, CHG = 2, CHH = 0, CxG = 0, CX = 0)[extract.context],
  highlight.positions = c(),
  ...,
  verbose = TRUE
)

Arguments

bam	BAM file location string OR preprocessed output of preprocessBam function. Read more about BAM file requirements and BAM preprocessing at preprocessBam.
bed	Browser Extensible Data (BED) file location string OR object of class GRanges holding genomic coordinates for regions of interest. It is used to match sequencing reads to the genomic regions for pattern extraction. The style of seqlevels of BED file/object must match the style of seqlevels of the BAM file/object used. The BED/GRanges rows are not sorted internally. As of now, the strand information is ignored and patterns matching both strands are extracted.
bed.row	single non-negative integer specifying what 'bed' region should be included in the output (default: 1).
zero.based.bed	boolean defining if BED coordinates are zero based (default: FALSE).
match.min.overlap	integer for the smallest overlap between read's and BED/GRanges start or end positions during matching of capture-based NGS reads (default: 1).
extract.context	string defining cytosine methylation context used to report: "CG" (the default) – CpG cytosines (called as zZ) "CHG" – CHG cytosines (xX) "CHH" – CHH cytosines (hH) "CxG" – CG and CHG cytosines (zZxX) "CX" – all cytosines
min.context.freq	real number in the range [0;1] (default: 0.01). Genomic positions that are covered by smaller fraction of patterns (e.g., with erroneous context) won't be included in the report.
clip.patterns	boolean if patterns should not extend over the edge of 'bed' region (default: FALSE).
strand.offset	single non-negative integer specifying the offset of bases at the reverse (-) strand compared to the forward (+) strand. Allows to "merge" genomic positions when methylation is symmetric (in CG and CHG contexts). By default, equals 1 for 'extract.context'=="CG", 2 for "CHG", or 0 otherwise.
highlight.positions	integer vector with genomic positions of bases to include in every overlapping pattern. Allows to visualize the distribution of single-nucleotide variations (SNVs) among methylation patterns. 'highlight.positions' takes precedence if any of these positions overlap with within-the-context positions of methylation pattern.
...	other parameters to pass to the preprocessBam function. Options have no effect if preprocessed BAM data was supplied as an input.
verbose	boolean to report progress and timings (default: TRUE).

Details

The function matches reads (for paired-end sequencing alignment files - read pairs as a single entity) to the genomic region provided in a BED file/GRanges object, extracts methylation statuses of bases within those reads, and returns a data frame which can be used for further analysis and/or plotting of DNA methylation patterns by plotPatterns function.

Value

data.table object containing per-read (pair) base methylation information for the genomic region of interest. The report columns are:

• seqnames – read (pair) reference sequence name

• strand – read (pair) strand

• start – start of the read (pair)

• end – end of the read (pair)

• nbase – number of within-the-context bases for this read (pair)

• beta – beta value of this read (pair), calculated as a ratio of the number of methylated within-the-context bases to the total number of within-the-context bases

• pattern – hex representation of 64-bit FNV-1a hash calculated for all reported base positions and bases in this read (pair). This hash value depends only on included genomic positions and their methylation call string chars (hHxXzZ) or nucleotides (ACGT, for highlighted bases only), thus it is expected to be unique for every methylation pattern, although equal for identical methylation patterns independently on read (pair) start, end, or strand (when correct 'strand.offset' is given)

• ... – columns for each genomic position that hold corresponding methylation call string char, or NA if position is not present in the read (pair)

See Also

plotPatterns for pretty plotting of the output, preprocessBam for preloading BAM data, generateCytosineReport for methylation statistics at the level of individual cytosines, generateBedReport for genomic region-based statistics, generateVcfReport for evaluating epiallele-SNV associations, generateBedEcdf for analysing the distribution of per-read beta values, and 'epialleleR' vignettes for the description of usage and sample data.

Examples

  # amplicon data
  amplicon.bam <- system.file("extdata", "amplicon010meth.bam",
                              package="epialleleR")
  amplicon.bed <- system.file("extdata", "amplicon.bed",
                              package="epialleleR")
  
  # extract patterns
  patterns <- extractPatterns(bam=amplicon.bam, bed=amplicon.bed, bed.row=3)
  
  # and then plot them
  plotPatterns(patterns)
  

BBCG/epialleleR documentation built on Nov. 4, 2024, 12:56 p.m.