How Do I Demultiplex Off-instrument?

If you have the unfiltered_fastq files, you can demultiplex your sequencing data using stand-alone demultiplexing software packages.

Choose one of these two packages:

• Use the Singular-Demux software if you want fine level control over the various options for demultiplexing your files. The software has additional functionalities for working with your Instrument FASTQ files.

• Use the Singular-Demux Folder Level Nextflow Workflow if you want an easy way to demultiplex your unfiltered_fastq data directory with your sample sheet using mostly default options.

Perform off-instrument demultiplexing the following way:

1. Correct the sample sheet.

2. Download and use one of these two packages to demultiplex off-instrument:

• Singular-Demux software, available at https://github.com/Singular-Genomics/singular-demux.

• Singular-Demux Folder Level Nextflow Workflow, available at https://github.com/Singular-Genomics/sgdemux_folder_nf2.

3. Follow the instructions available in the repository.

See also the Technical Bulletin Guide to Working with G4 Sequencing Data.

11/29/2023