Microfluidic approaches to next- generation sequencing library preparation: innovations, clinical integration and...

Next-generation sequencing (NGS) has revolutionized genomics by enabling high-throughput, cost-effective analysis of nucleic acids for both research and clinical applications. Library preparation remains a bottleneck in sequencing due to high reagent use, labor-intensive workflows, long hands-on times, and susceptibility to human error, despite advancements in sequencing technology.

This review emphasizes the latest advancements in microfluidics-based next-generation sequencing (NGS) library preparation, focusing on four specific platform types: continuous-flow systems, droplet microfluidics, digital microfluidics (DMF), and integrated lab-on-chip devices. These innovative technologies significantly reduce reagent consumption by 10 to 200 times, facilitate the creation of high-quality libraries from picogram-scale DNA or even single cells, and automate intricate workflows within enclosed systems. The applications of these technologies in liquid biopsy, pathogen detection, prenatal testing, and epigenomic profiling showcase considerable clinical promise. Challenges such as fabrication complexity, lack of standardization, biofouling, and high instrument costs hinder widespread adoption, particularly in resource-limited settings. However, advancements in AI, thermal control, long-read sequencing, and portable microfluidics are helping to overcome these issues.

Microfluidic library preparation has the potential to transform NGS workflows by enhancing automation, reproducibility, and accessibility, thereby facilitating broader applications of genomic sequencing in research, clinical diagnostics, and point-of-care settings.