SangerContig-class: SangerContig

In sangeranalyseR: sangeranalyseR: a suite of functions for the analysis of Sanger sequence data in R

Description

An S4 class containing forward and reverse SangerRead lists and alignment, consensus read results which corresponds to a contig in Sanger sequencing.

Slots

inputSource

The input source of the raw file. It must be "ABIF" or "FASTA". The default value is "ABIF".

fastaFileName

If inputSource is "FASTA", then this value has to be the FASTA file; if inputSource is "ABIF", then this value is NULL by default.

namesConversionCSV

The file path to the CSV file that provides read names that follow the naming regulation. If inputSource is "FASTA", then users need to prepare the csv file or make sure the original names inside FASTA file are valid; if inputSource is "ABIF", then this value is NULL by default.

parentDirectory

If inputSource is "ABIF", then this value is the path of the parent directory storing all reads in ABIF format you wish to analyse and cannot be NULL. In SangerContig, all reads must be in the first layer in this directory. If inputSource is "FASTA", then this value is NULL by default.

contigName

The contig name of all the reads in parentDirectory.

suffixForwardRegExp

The suffix of the filenames for forward reads in regular expression, i.e. reads that do not need to be reverse-complemented. For forward reads, it should be "_F.ab1".

suffixReverseRegExp

The suffix of the filenames for reverse reads in regular expression, i.e. reads that need to be reverse-complemented. For revcerse reads, it should be "_R.ab1".

geneticCode

Named character vector in the same format as GENETIC_CODE (the default), which represents the standard genetic code. This is the code with which the function will attempt to translate your DNA sequences. You can get an appropriate vector with the getGeneticCode() function. The default is the standard code.

forwardReadList

The list of SangerRead S4 instances which are all forward reads.

reverseReadList

The list of SangerRead S4 instances which are all reverse reads.

trimmingMethodSC

The read trimming method for all SangerRead S4 instances in SangerContig. The value must be "M1" (the default) or 'M2'. All SangerRead must have the same trimming method.

minReadsNum

The minimum number of reads required to make a consensus sequence, must be 2 or more. The default value is 2.

minReadLength

Reads shorter than this will not be included in the readset. The default 20 means that all reads with length of 20 or more will be included. Note that this is the length of a read after it has been trimmed.

refAminoAcidSeq

An amino acid reference sequence supplied as a string or an AAString object. If your sequences are protein-coding DNA seuqences, and you want to have frameshifts automatically detected and corrected, supply a reference amino acid sequence via this argument. If this argument is supplied, the sequences are then kept in frame for the alignment step. Fwd sequences are assumed to come from the sense (i.e. coding, or "+") strand. The default value is "".

minFractionCall

Minimum fraction of the sequences required to call a consensus sequence for SangerContig at any given position (see the ConsensusSequence() function from DECIPHER for more information). Defaults to 0.75 implying that 3/4 of all reads must be present in order to call a consensus.

maxFractionLost

Numeric giving the maximum fraction of sequence information that can be lost in the consensus sequence for SangerContig (see the ConsensusSequence() function from DECIPHER for more information). Defaults to 0.5, implying that each consensus base can ignore at most 50 percent of the information at a given position.

acceptStopCodons

The logical value TRUE or FALSE. TRUE (the defualt): keep all reads, regardless of whether they have stop codons; FALSE: reject reads with stop codons. If FALSE is selected, then the number of stop codons is calculated after attempting to correct frameshift mutations (if applicable).

readingFrame

1, 2, or 3. Only used if accept.stop.codons == FALSE. This specifies the reading frame that is used to determine stop codons. If you use a refAminoAcidSeq, then the frame should always be 1, since all reads will be shifted to frame 1 during frameshift correction. Otherwise, you should select the appropriate reading frame.

contigSeq

The consensus read of all SangerRead S4 instances in DNAString object.

alignment

The alignment of all SangerRead S4 instances with the called consensus sequence in DNAStringSet object. Users can use BrowseSeqs() to view the alignment.

differencesDF

A data frame of the number of pairwise differences between each read and the consensus sequence, as well as the number of bases in each input read that did not contribute to the consensus sequence. It can assist in detecting incorrect reads, or reads with a lot of errors.

distanceMatrix

A distance matrix of genetic distances (corrected with the JC model) between all of the input reads.

dendrogram

A list storing cluster groups in a data frame and a dendrogram object depicting the distance.matrix. Users can use plot() to see the dendrogram.

indelsDF

If users specified a reference sequence via refAminoAcidSeq, then this will be a data frame describing the number of indels and deletions that were made to each of the input reads in order to correct frameshift mutations.

stopCodonsDF

If users specified a reference sequence via refAminoAcidSeq, then this will be a data frame describing the number of stop codons in each read.

secondaryPeakDF

A data frame with one row for each column in the alignment that contained more than one secondary peak. The data frame has three columns: the column number of the alignment; the number of secondary peaks in that column; and the bases (with IUPAC ambiguity codes representing secondary peak calls) in that column represented as a string.

Author(s)

Kuan-Hao Chao

Examples

## Input From ABIF file format (Regex)
rawDataDir <- system.file("extdata", package = "sangeranalyseR")
parentDir <- file.path(rawDataDir, "Allolobophora_chlorotica", "RBNII")
contigName <- "Achl_RBNII384-13"
suffixForwardRegExp <- "_[0-9]*_F.ab1"
suffixReverseRegExp <- "_[0-9]*_R.ab1"
sangerContig <- new("SangerContig",
                     inputSource           = "ABIF",
                     parentDirectory       = parentDir,
                     contigName            = contigName,
                     suffixForwardRegExp   = suffixForwardRegExp,
                     suffixReverseRegExp   = suffixReverseRegExp,
                     refAminoAcidSeq = "SRQWLFSTNHKDIGTLYFIFGAWAGMVGTSLSILIRAELGHPGALIGDDQIYNVIVTAHAFIMIFFMVMPIMIGGFGNWLVPLMLGAPDMAFPRMNNMSFWLLPPALSLLLVSSMVENGAGTGWTVYPPLSAGIAHGGASVDLAIFSLHLAGISSILGAVNFITTVINMRSTGISLDRMPLFVWSVVITALLLLLSLPVLAGAITMLLTDRNLNTSFFDPAGGGDPILYQHLFWFFGHPEVYILILPGFGMISHIISQESGKKETFGSLGMIYAMLAIGLLGFIVWAHHMFTVGMDVDTRAYFTSATMIIAVPTGIKIFSWLATLHGTQLSYSPAILWALGFVFLFTVGGLTGVVLANSSVDIILHDTYYVVAHFHYVLSMGAVFAIMAGFIHWYPLFTGLTLNNKWLKSHFIIMFIGVNLTFFPQHFLGLAGMPRRYSDYPDAYTTWNIVSTIGSTISLLGILFFFFIIWESLVSQRQVIYPIQLNSSIEWYQNTPPAEHSYSELPLLTN",
                     TrimmingMethod        = "M1",
                     M1TrimmingCutoff      = 0.0001,
                     M2CutoffQualityScore  = NULL,
                     M2SlidingWindowSize   = NULL,
                     baseNumPerRow         = 100,
                     heightPerRow          = 200,
                     signalRatioCutoff     = 0.33,
                     showTrimmed           = TRUE,
                     processorsNum         = 2)

## Input From ABIF file format (Csv three column method)
rawDataDir <- system.file("extdata", package = "sangeranalyseR")
parentDir <- file.path(rawDataDir, "Allolobophora_chlorotica", "RBNII")
namesConversionCSV <- file.path(rawDataDir, "ab1", "SangerContig", "names_conversion_2.csv")
sangerContig <- new("SangerContig",
                     inputSource           = "ABIF",
                     parentDirectory       = parentDir,
                     namesConversionCSV    = namesConversionCSV,
                     refAminoAcidSeq = "SRQWLFSTNHKDIGTLYFIFGAWAGMVGTSLSILIRAELGHPGALIGDDQIYNVIVTAHAFIMIFFMVMPIMIGGFGNWLVPLMLGAPDMAFPRMNNMSFWLLPPALSLLLVSSMVENGAGTGWTVYPPLSAGIAHGGASVDLAIFSLHLAGISSILGAVNFITTVINMRSTGISLDRMPLFVWSVVITALLLLLSLPVLAGAITMLLTDRNLNTSFFDPAGGGDPILYQHLFWFFGHPEVYILILPGFGMISHIISQESGKKETFGSLGMIYAMLAIGLLGFIVWAHHMFTVGMDVDTRAYFTSATMIIAVPTGIKIFSWLATLHGTQLSYSPAILWALGFVFLFTVGGLTGVVLANSSVDIILHDTYYVVAHFHYVLSMGAVFAIMAGFIHWYPLFTGLTLNNKWLKSHFIIMFIGVNLTFFPQHFLGLAGMPRRYSDYPDAYTTWNIVSTIGSTISLLGILFFFFIIWESLVSQRQVIYPIQLNSSIEWYQNTPPAEHSYSELPLLTN",
                     TrimmingMethod        = "M1",
                     M1TrimmingCutoff      = 0.0001,
                     M2CutoffQualityScore  = NULL,
                     M2SlidingWindowSize   = NULL,
                     baseNumPerRow         = 100,
                     heightPerRow          = 200,
                     signalRatioCutoff     = 0.33,
                     showTrimmed           = TRUE,
                     processorsNum         = 2)


## Input From FASTA file format (No Csv - Regex)
rawDataDir <- system.file("extdata", package = "sangeranalyseR")
fastaFN <- file.path(rawDataDir, "fasta",
                     "SangerContig", "Achl_ACHLO006-09.fa")
contigName <- "Achl_ACHLO006-09"
suffixForwardRegExpFa <- "_[0-9]*_F$"
suffixReverseRegExpFa <- "_[0-9]*_R$"
sangerContigFa <- new("SangerContig",
                      inputSource           = "FASTA",
                      fastaFileName         = fastaFN,
                      contigName            = contigName,
                      suffixForwardRegExp   = suffixForwardRegExpFa,
                      suffixReverseRegExp   = suffixReverseRegExpFa,
                      refAminoAcidSeq       = "SRQWLFSTNHKDIGTLYFIFGAWAGMVGTSLSILIRAELGHPGALIGDDQIYNVIVTAHAFIMIFFMVMPIMIGGFGNWLVPLMLGAPDMAFPRMNNMSFWLLPPALSLLLVSSMVENGAGTGWTVYPPLSAGIAHGGASVDLAIFSLHLAGISSILGAVNFITTVINMRSTGISLDRMPLFVWSVVITALLLLLSLPVLAGAITMLLTDRNLNTSFFDPAGGGDPILYQHLFWFFGHPEVYILILPGFGMISHIISQESGKKETFGSLGMIYAMLAIGLLGFIVWAHHMFTVGMDVDTRAYFTSATMIIAVPTGIKIFSWLATLHGTQLSYSPAILWALGFVFLFTVGGLTGVVLANSSVDIILHDTYYVVAHFHYVLSMGAVFAIMAGFIHWYPLFTGLTLNNKWLKSHFIIMFIGVNLTFFPQHFLGLAGMPRRYSDYPDAYTTWNIVSTIGSTISLLGILFFFFIIWESLVSQRQVIYPIQLNSSIEWYQNTPPAEHSYSELPLLTN",
                      processorsNum         = 2)

## Input From FASTA file format (Csv - Csv three column method)
rawDataDir <- system.file("extdata", package = "sangeranalyseR")
fastaFN <- file.path(rawDataDir, "fasta",
                     "SangerContig", "Achl_ACHLO006-09.fa")
namesConversionCSV <- file.path(rawDataDir, "fasta", "SangerContig", "names_conversion_1.csv")
sangerContigFa <- new("SangerContig",
                      inputSource           = "FASTA",
                      fastaFileName         = fastaFN,
                      namesConversionCSV    = namesConversionCSV,
                      refAminoAcidSeq       = "SRQWLFSTNHKDIGTLYFIFGAWAGMVGTSLSILIRAELGHPGALIGDDQIYNVIVTAHAFIMIFFMVMPIMIGGFGNWLVPLMLGAPDMAFPRMNNMSFWLLPPALSLLLVSSMVENGAGTGWTVYPPLSAGIAHGGASVDLAIFSLHLAGISSILGAVNFITTVINMRSTGISLDRMPLFVWSVVITALLLLLSLPVLAGAITMLLTDRNLNTSFFDPAGGGDPILYQHLFWFFGHPEVYILILPGFGMISHIISQESGKKETFGSLGMIYAMLAIGLLGFIVWAHHMFTVGMDVDTRAYFTSATMIIAVPTGIKIFSWLATLHGTQLSYSPAILWALGFVFLFTVGGLTGVVLANSSVDIILHDTYYVVAHFHYVLSMGAVFAIMAGFIHWYPLFTGLTLNNKWLKSHFIIMFIGVNLTFFPQHFLGLAGMPRRYSDYPDAYTTWNIVSTIGSTISLLGILFFFFIIWESLVSQRQVIYPIQLNSSIEWYQNTPPAEHSYSELPLLTN",
                      processorsNum         = 2)

sangeranalyseR documentation built on Nov. 8, 2020, 5:59 p.m.