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Related Experiment Videos

scSPLAT, a scalable plate-based protocol for single cell WGBS library preparation.

Amanda Raine1, Anders Lundmark2, Alva Annett2

  • 1Science for Life Laboratory, Department of Medical Sciences, Uppsala University, Uppsala, Sweden. Amanda.Raine@medsci.uu.se.

Scientific Reports
|April 7, 2022
PubMed
Summary

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We developed single-cell Splinted Ligation Adapter Tagging (scSPLAT), a cost-effective method for DNA methylation analysis. This technique enhances scalability and throughput for whole-genome bisulfite sequencing (scWGBS) in single cells.

Area of Science:

  • Epigenetics
  • Genomics
  • Molecular Biology

Background:

  • DNA methylation is a crucial epigenetic modification regulating gene expression, development, and genome stability.
  • Single-cell whole-genome bisulfite sequencing (scWGBS) enables base-resolution interrogation of 5-methylcytosine (5mC) in individual cells.
  • Existing scWGBS methods face challenges in cost-efficiency and scalability.

Purpose of the Study:

  • To introduce a novel, scalable, and cost-effective library preparation method for scWGBS.
  • To improve the throughput of single-cell DNA methylation analysis.
  • To validate the accuracy and robustness of the new method.

Main Methods:

  • Development of single-cell Splinted Ligation Adapter Tagging (scSPLAT) for scWGBS library preparation.

Related Experiment Videos

  • Implementation of a pooling strategy to enhance sample preparation scale and throughput.
  • Generation of scWGBS data from 225 single K562 cells and 309 single liver nuclei.

    • Main Results:

    • scSPLAT demonstrates accuracy and robustness in DNA methylation profiling at single-cell resolution.
    • The method facilitates higher scale and throughput compared to previous scWGBS strategies.
    • Comparative analysis validates scSPLAT's performance against existing scWGBS techniques.

    Conclusions:

    • scSPLAT represents a significant advancement in scWGBS library preparation, addressing limitations in cost and yield.
    • The method enables large-scale, high-throughput DNA methylation analysis in single cells.
    • scSPLAT is a valuable tool for studying epigenetic heterogeneity and dynamics in biological systems.