sabre - A barcode demultiplexing and trimming tool for FastQ files

About

Next-generation sequencing can currently produce hundreds of millions of reads per lane of sample and that number increases at a dizzying rate. Barcoding individual sequences for multiple lines or multiple species is a cost-efficient method to sequence and analyze a broad range of data.

Sabre is a tool that will demultiplex barcoded reads into separate files. It will work on both single-end and paired-end data in fastq format. It simply compares the provided barcodes with each read and separates the read into its appropriate barcode file, after stripping the barcode from the read (and also stripping the quality values of the barcode bases). If a read does not have a recognized barcode, then it is put into the unknown file.
Sabre also has an option (-m) to allow mismatches of the barcodes.

Sabre also supports gzipped file inputs. Also, since sabre does not use the quality values in any way, it can be used on fasta data that is converted to fastq by creating fake quality values.

Finally, after demultiplexing, sabre outputs a summary of how many records went into each barcode file.

Requirements

Sabre requires a C compiler; GCC or clang are recommended. Sabre relies on Heng Li's kseq.h, which is bundled with the source.

Sabre also requires Zlib, which can be obtained at http://www.zlib.net/.

Building and Installing Sabre

To build Sabre, enter:

make

Then, copy or move "sabre" to a directory in your $PATH.

Usage

Sabre has two modes to work with both paired-end and single-end reads: sabre se and sabre pe.

Running sabre by itself will print the help:

sabre

Running sabre with either the "se" or "pe" commands will give help specific to those commands:

sabre se
sabre pe

Sabre Single End (sabre se)

sabre se takes an input fastq file and an input barcode data file and outputs the reads demultiplexed into separate files using the file names from the data file. The barcodes will be stripped from the reads and the quality values of the barcode bases will also be removed. Any reads with unknown barcodes get put into the "unknown" file specified on the command line. The -m option allows for mismatches in the barcodes.

Barcode data file format for single end

barcode1 barcode1_output_file.fastq
barcode2 barcode2_output_file.fastq
etc...

Be aware that if you do not format the barcode data file correctly, sabre will not work properly.

Example

sabre se -f input_file.fastq -b barcode_data.txt -u unknown_barcode.fastq
sabre se -m 1 -f input_file.fastq -b barcode_data.txt -u unknown_barcode.fastq

Sabre Paired End (sabre pe)

sabre pe takes two paired-end files and a barcode data file as input and outputs the reads demultiplexed into separate paired-end files using the file names from the data file. The barcodes will be stripped from the reads and the quality values of the barcode bases will also be removed. Any reads with unknown barcodes get put into the "unknown" files specified on the command line. It also has an option (-c) to remove barcodes from both files.
Using this option means that if sabre finds a barcode in the first file, it assumes the paired read in the other file has the same barcode and will strip it (along with the quality values).
The -m option allows for mismatches in the barcodes.

Barcode data file format for paired end

barcode1 barcode1_output_file1.fastq barcode1_output_file2.fastq
barcode2 barcode2_output_file1.fastq barcode2_output_file2.fastq
etc...

Be aware that if you do not format the barcode data file correctly, sabre will not work properly.

Examples

sabre pe -f input_file1.fastq -r input_file2.fastq -b barcode_data.txt \
-u unknown_barcode1.fastq -w unknown_barcode1.fastq

sabre pe -c -f input_file1.fastq -r input_file2.fastq -b barcode_data.txt \
-u unknown_barcode1.fastq -w unknown_barcode1.fastq

sabre pe -m 2 -f input_file1.fastq -r input_file2.fastq -b barcode_data.txt \
-u unknown_barcode1.fastq -w unknown_barcode1.fastq