SASI-Seq: sample assurance Spike-Ins, and highly differentiating 384 barcoding for Illumina sequencing

Area of Science:

• Genomics
• Molecular Biology
• Bioinformatics

Background:

• Next-generation sequencing (NGS) is susceptible to sample mix-ups and cross-contamination.
• These errors can compromise the accuracy and reliability of sequencing results.
• Ensuring sample integrity is crucial for valid experimental outcomes.

Purpose of the Study:

• To develop a method for verifying sample origin and detecting contamination in NGS.
• To introduce SASI-Seq as a tool for sample assurance.
• To quantify cross-contamination levels in sequencing workflows.

Main Methods:

• SASI-Seq involves adding uniquely barcoded DNA fragments to samples before sequencing.
• These fragments are designed to remain associated with the original sample throughout library preparation.
• A set of 384 eleven-base Illumina barcodes with high error tolerance was developed.

Main Results:

• SASI-Seq fragments were detected even after stringent size selection and exome enrichment.
• The method successfully identified sample mix-ups and cross-contamination.
• The developed barcodes allow for single-error correction, minimizing misallocation.

Conclusions:

• SASI-Seq is a simple, cost-effective, and adaptable solution for sample assurance in NGS.
• The technology effectively deconvolutes sample mix-ups and quantifies cross-contamination.
• SASI-Seq enhances the reliability of data generated from various NGS applications.