Barbell

Barbell demultiplexes Oxford Nanopore sequencing data by detecting all barcode configurations including complex barcode attachments, reducing trimming errors and barcode bleeding compared to existing demultiplexers.

Key Features:

• Pattern-aware barcode detection: Detects all barcode configurations including multiple barcodes at the same read end, identical barcodes from the same sample, and different barcodes introduced after pooling.
• Error reduction: Reduces trimming errors by three orders of magnitude compared to existing demultiplexers such as Dorado.
• Barcode bleeding minimization: Prevents incorrect selection of outer barcodes when different barcodes are present at the same read end after pooling.
• Custom experimental setup support: Supports shorter barcodes, dual-end barcodes, and custom flank sequences.

Scientific Applications:

• Diagnostic sample sequencing: Accurate demultiplexing of rapid barcoding experiments on diagnostic samples where 17% of reads may contain multiple barcodes or other artifacts.
• Downstream analysis improvement: Eliminates contamination with adapter fragments from approximately 7% of reads that are partially trimmed by existing demultiplexers.
• Custom barcoding workflows: Enables flexible demultiplexing for non-standard experimental designs with modified barcode lengths or flank sequences.

Methodology:

Barbell analyzes Oxford Nanopore reads to detect all barcode configurations through pattern-aware detection of single and multiple barcode attachments at read ends.