A plate-based single-cell ATAC-seq workflow for fast and robust profiling of chromatin accessibility

Area of Science:

• Molecular Biology
• Genomics
• Cell Biology

Background:

• Single-cell chromatin accessibility profiling is crucial for understanding cellular heterogeneity.
• Existing methods can be time-consuming and complex.
• Investigating cell-to-cell variation requires efficient and robust techniques.

Purpose of the Study:

• To develop a fast and robust protocol for single-cell chromatin accessibility profiling.
• To enable detailed characterization of cell types and their variations.
• To provide a method compatible with standard molecular biology techniques.

Main Methods:

• A streamlined single-cell assay for transposase-accessible chromatin using sequencing (ATAC-seq).
• Combines bulk Tn5 enzyme tagging with flow cytometry for single-cell isolation.
• Reagents for library preparation are added post-sorting in the same plate.

Main Results:

• Generates high-complexity data with excellent signal-to-noise ratio.
• Protocol completion within 1-2 days with high throughput (hundreds to thousands of nuclei).
• Data is processable via a provided computational pipeline.

Conclusions:

• The developed protocol offers an efficient and accessible method for single-cell ATAC-seq.
• Suitable for basic molecular biology labs with flow cytometry capabilities.
• Facilitates in-depth characterization of cellular heterogeneity.