All-in-one sequencing: an improved library preparation method for cost-effective and high-throughput next-generation...

Area of Science:

• Genomics
• Molecular Biology
• Bioinformatics

Background:

• Next-generation sequencing (NGS) is crucial in biological research, but library preparation remains inefficient.
• Time-consuming steps like size-selection and quantification create bottlenecks, especially for large sample cohorts.

Purpose of the Study:

• To develop a more efficient library preparation method for NGS.
• To reduce labor and costs associated with processing large numbers of samples.

Main Methods:

• Introduced the all-in-one sequencing (AIO-seq) method, pooling up to 116 libraries in a single tube.
• Libraries are pooled based on expected data yields and quantified target region concentrations.
• Simplified protocol demonstrated for low-coverage sequencing data, like recombinant inbred lines (RILs), by pooling post-PCR.

Main Results:

• AIO-seq successfully applied to whole genome sequencing and RNA-seq libraries.
• Demonstrated applicability to various NGS library types, including ChIP-seq, ATAC-seq, and Hi-C.
• Simplified AIO-seq protocol validated for specific genetic population samples.

Conclusions:

• AIO-seq is a labor-saving and cost-effective method for NGS library preparation.
• The method is highly suitable for projects involving large sample cohorts, such as plant breeding and population genetics research.