Construction of Strand-seq libraries in open nanoliter arrays

Area of Science:

• Genomics
• Molecular Biology
• Biotechnology

Background:

• Single-cell Strand-seq is a technique for obtaining directional genomic information.
• This information is valuable for studying DNA repair, genome assembly, and mapping structural variations.
• Current methods can be complex and costly.

Purpose of the Study:

• To develop a simplified and cost-effective Strand-seq library preparation protocol.
• To increase the throughput of single-cell genomic analysis using Strand-seq.

Main Methods:

• A nanoliter-volume, one-pot (OP) protocol was developed for Strand-seq library preparation.
• Reagents were added cumulatively, eliminating DNA purification steps.
• A thermolabile protease was used for enzyme inactivation.

Main Results:

• The OP-Strand-seq protocol successfully prepared libraries from single cells.
• These libraries captured 10%-25% of the genome per cell.
• The protocol demonstrated reduced costs and increased throughput compared to existing methods.

Conclusions:

• The one-pot Strand-seq protocol is an efficient method for single-cell genomic analysis.
• This streamlined approach lowers costs and enhances throughput for studying DNA repair and structural variations.