Type: Package
Title: Assembling Long Gene Copies from Short Read Data
Version: 1.2.0
Author: Lei Yang
Maintainer: Lei Yang <leiyangslu@gmail.com>
Description: Assembles two or more gene copies from short-read Next-Generation Sequencing data. Works best when there are only two gene copies and read length >=250 base pairs. High and relatively even coverage are important.
License: GPL-2
URL: https://github.com/LeiYang-Fish/copyseparator
BugReports: https://github.com/LeiYang-Fish/copyseparator/issues
Depends: R (≥ 3.5.0)
Encoding: UTF-8
Imports: ape, seqinr, stringr, kmer, DECIPHER, beepr, Biostrings, grDevices, doParallel, foreach, parallel
RoxygenNote: 7.2.1
Suggests: knitr, rmarkdown, testthat (≥ 3.0.0)
VignetteBuilder: knitr
NeedsCompilation: no
Packaged: 2022-11-25 00:14:04 UTC; lyang1
Repository: CRAN
Date/Publication: 2022-11-25 00:30:02 UTC

copy_assemble

Description

Assembles a small number of overlapping DNA sequences into their respective gene copies.

Usage

copy_assemble(filename, copy_number, verbose = 1)

Arguments

filename

A fasta alignment of a small number of overlapping DNA sequences (results from "copy_separate") covering the entire length of the target gene. Check the alignment carefully before proceeding.

copy_number

An integer (e.g. 2,3, or 4) giving the anticipated number of gene copies. Must be the same value as used for "copy_separate".

verbose

Turn on (verbose=1; default) or turn off (verbose=0) the output.

Value

A fasta alignment of the anticipated number of full-length gene copies.

Examples

## Not run: 
copy_assemble("inst/extdata/combined_con.fasta",2,1)

## End(Not run)


copy_detect

Description

Separates two or more gene copies from a single subset of short reads.

Usage

copy_detect(filename, copy_number, verbose = 1)

Arguments

filename

A fasta file contains short reads from a single subset generated by "subset_downsize".

copy_number

An integer (e.g. 2,3, or 4) giving the anticipated number of gene copies in the input file.

verbose

Turn on (verbose=1; default) or turn off (verbose=0) the output.

Value

A fasta alignment of the anticipated number of gene copies.

Examples

## Not run: 
copy_detect("inst/extdata/toysubset.fasta",2,1)

## End(Not run)


copy_separate

Description

Separates two or more gene copies from short-read Next-Generation Sequencing data into a small number of overlapping DNA sequences.

Usage

copy_separate(
  filename,
  copy_number,
  read_length,
  overlap = 225,
  rare_read = 10,
  verbose = 1
)

Arguments

filename

A fasta file contains thousands of short reads that have been mapped to a reference. The reference and reads that are not directly mapped to the reference need to be removed after mapping.

copy_number

An integer (e.g. 2,3, or 4) giving the anticipated number of gene copies in the input file.

read_length

An integer (e.g. 250, or 300) giving the read length of your Next-generation Sequencing data. This method is designed for read length >=250bp.

overlap

An integer describing number of base pairs of overlap between adjacent subsets. More overlap means more subsets. Default 225.

rare_read

A positive integer. During clustering analyses, clusters with less than this number of reads will be ignored. Default 10.

verbose

Turn on (verbose=1; default) or turn off (verbose=0) the output.

Value

A fasta alignment of a small number of overlapping DNA sequences covering the entire length of the target gene. Gene copies can be assembled by reordering the alignment manually or use the function "copy_assemble".

Examples

## Not run: 
copy_separate("inst/extdata/toydata.fasta",2,300,225,10,1)

## End(Not run)


copy_validate

Description

A tool to help identify incorrectly assembled chimeric sequences.

Usage

copy_validate(filename, copy_number, read_length, verbose = 1)

Arguments

filename

A DNA alignment in fasta format that contains sequences of two or more gene copies (e.g. results from "copy_assemble").

copy_number

An integer (e.g. 2,3, or 4) giving the number of gene copies in the input file.

read_length

An integer (e.g. 250, or 300) giving the read length of your Next-generation Sequencing data.

verbose

Turn on (verbose=1; default) or turn off (verbose=0) the output.

Value

A histogram in pdf format showing the relationships between the physical distance between neighboring variable sites and read length.

Examples

## Not run: 
copy_validate("inst/extdata/Final_two_copies.fasta",2,300,1)

## End(Not run)


sep_assem

Description

Separates two or more gene copies from short-read Next-Generation Sequencing data into a small number of overlapping DNA sequences and assemble them into their respective gene copies.

Usage

sep_assem(
  copy_number,
  read_length,
  overlap = 225,
  rare_read = 10,
  core_number = 1,
  verbose = 1
)

Arguments

copy_number

An integer (e.g. 2,3, or 4) giving the anticipated number of gene copies in the input file.

read_length

An integer (e.g. 250, or 300) giving the read length of your Next-generation Sequencing data. This method is designed for read length >=250bp.

overlap

An integer describing number of base pairs of overlap between adjacent subsets. More overlap means more subsets. Default 225.

rare_read

A positive integer. During clustering analyses, clusters with less than this number of reads will be ignored. Default 10.

core_number

An integer describing number of cores to use.

verbose

Turn on (verbose=1; default) or turn off (verbose=0) the output.

Value

A fasta alignment of the anticipated number of full-length gene copies.

Examples

## Not run: 
sep_assem(2,300,225,10,1,1) # all input fasta files in the working directory will be processed

## End(Not run)


subset_downsize

Description

Subdivides the imported read alignment into subsets and then downsizes each subset by deleting those sequences that have too many gaps or missing data.

Usage

subset_downsize(filename, read_length, overlap, verbose = 1)

Arguments

filename

A fasta file contains thousands of short reads that have been mapped to a reference. The reference and reads that are not directly mapped to the reference need to be removed after mapping.

read_length

An integer (e.g. 250, or 300) giving the read length of your Next-generation Sequencing data. This method is designed for read length >=250bp.

overlap

An integer describing number of base pairs of overlap between adjacent subsets. More overlap means more subsets.

verbose

Turn on (verbose=1; default) or turn off (verbose=0) the output.

Value

A number of overlapping subsets (before and after downsizing) of the input alignment.

Examples

## Not run: 
subset_downsize("inst/extdata/toydata.fasta",300,225,1)

## End(Not run)