Consistent ultra-long DNA sequencing with automated slow pipetting.

Area of Science:

• Genomics
• Molecular Biology
• Bioinformatics

Background:

• Oxford Nanopore Technologies enables long-read sequencing, crucial for resolving complex genomic regions.
• Preserving long DNA molecules during library preparation is essential for achieving significant read lengths.
• Manual slow pipetting is time-consuming and prone to shearing, impacting read length consistency.

Purpose of the Study:

• To develop an automated, low-cost instrument for precise, slow pipetting in long-read library preparation.
• To improve the preservation of long DNA template molecules during Oxford Nanopore library preparation.

Main Methods:

• Designed and 3D-printed SNAILS (Slow Nucleic Acid Instrument for Long Sequences).
• Automated slow pipetting of reagents for Oxford Nanopore library preparation.
• Compared read length preservation and average read length against manual pipetting.

Main Results:

• SNAILS preserved a higher proportion of DNA reads exceeding 100 kilobases.
• Libraries prepared with SNAILS showed an increased average read length compared to manual methods.
• The instrument demonstrated consistency in preserving long DNA template molecules.

Conclusions:

• SNAILS offers a low-cost, deployable solution for enhancing long-read sequencing projects.
• Automation of slow pipetting by SNAILS improves consistency and throughput for Nanopore sequencing.
• This instrument facilitates more reliable generation of long DNA reads for genomic analysis.