Sample enrichment for single-nucleus sequencing using concanavalin A-conjugated magnetic beads

Area of Science:

• Molecular Biology
• Genomics
• Cell Biology

Background:

• Single-cell and single-nucleus sequencing are powerful tools for cellular heterogeneity analysis.
• Sample dilution during isolation and difficulty in reconcentration via centrifugation pose challenges in these workflows.
• Efficient retrieval of specific cell populations is crucial for downstream genomic analysis.

Purpose of the Study:

• To develop and present an effective protocol for enriching dilute cell and nucleus samples.
• To improve the recovery rate of cells/nuclei after fluorescence-activated cell sorting (FACS).
• To integrate a sample enrichment step into existing single-cell/nucleus sequencing protocols, including 10x Genomics.

Main Methods:

• The protocol involves tissue dissection and nuclei isolation.
• Fluorescence-activated cell sorting (FACS) is employed for initial cell population selection.
• Concanavalin A-conjugated magnetic beads are utilized for efficient sample enrichment post-FACS.

Main Results:

• The developed protocol demonstrates efficient enrichment of cells and nuclei following FACS.
• The method overcomes challenges associated with sample dilution and poor recovery.
• The protocol seamlessly integrates with standard library preparation for 10x Genomics and other platforms.

Conclusions:

• This protocol provides a robust solution for enhancing cell/nucleus recovery in single-cell/nucleus sequencing.
• It facilitates more effective downstream analysis by ensuring higher sample yield.
• The method is adaptable and valuable for researchers utilizing FACS and single-cell genomics techniques.