Bead-linked transposomes enable a normalization-free workflow for NGS library preparation

Area of Science:

• Genomics
• Molecular Biology
• Next-Generation Sequencing

Background:

• Transposome-based DNA library preparation is crucial for sequencing.
• Current methods are sensitive to DNA input amount and quality.

Purpose of the Study:

• Introduce Nextera DNA Flex, a novel bead-based library preparation technology.
• Demonstrate its performance with diverse DNA inputs and challenging samples.

Main Methods:

• Utilized bead-conjugated transposomes for fixed DNA binding.
• Integrated DNA extraction protocol for direct sample input (blood, saliva).
• Tested performance with low DNA input (<1 ng), varying insert sizes, degraded FFPE samples, and diverse genomes.

Main Results:

• Nextera DNA Flex accommodates a wide range of DNA input, with yield saturation at 100 ng.
• Achieved uniform coverage in small genomes and improved coverage in large, complex genomes.
• Successfully prepared libraries from FFPE samples and amplicons (50 bp to 11 kb), with reduced efficiency for <250 bp amplicons.

Conclusions:

• Bead-based technology normalizes libraries, reducing the need for DNA quantitation.
• Offers robustness and flexibility for various DNA input types and amounts.
• Facilitates broad applications across genomic research fields.