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Related Experiment Video

Updated: Nov 15, 2025

Single-cell RNA Sequencing of Fluorescently Labeled Mouse Neurons Using Manual Sorting and Double In Vitro Transcription with Absolute Counts Sequencing DIVA-Seq
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ATAC-seq on Sorted Adult Mouse Neurons.

Erin A Clark1, Yasuyuki Shima1, Sacha Nelson1

  • 1Department of Biology and Program in Neuroscience, Brandeis University, Waltham, USA.

Bio-Protocol
|March 3, 2021
PubMed
Summary

Researchers developed a new method to prepare live neurons for the Assay for Transposase Accessible Chromatin (ATAC). This technique enables detailed study of transcription regulation and cell identity in the adult mammalian brain.

Keywords:
ATACChromatinNeuronTn5Transposase

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Area of Science:

  • Neurobiology
  • Genomics
  • Molecular Biology

Background:

  • Transcription regulation is crucial for cellular identity in development and adulthood.
  • Understanding cell-type-specific functions in the brain requires selective study of neuronal populations.
  • Limited cellular material often hinders molecular assays in neurobiology.

Purpose of the Study:

  • To describe a novel method for preparing live neurons from adult brain tissue.
  • To facilitate the application of genome-probing assays, such as ATAC, in neurobiology.
  • To overcome challenges in obtaining sufficient cellular input for molecular analyses in neurons.

Main Methods:

  • Development of a specialized protocol for dissociating and preparing live neurons from adult mammalian brain.
  • Adaptation of existing molecular assay methodologies for low-input samples.
  • Focus on preserving cellular integrity and biological information during sample preparation for the Assay for Transposase Accessible Chromatin (ATAC).

Main Results:

  • Successful preparation of live, viable neurons suitable for downstream molecular assays.
  • Demonstration of a method compatible with the Assay for Transposase Accessible Chromatin (ATAC).
  • Establishment of a technique to address the challenge of limited cellular input in neurobiological studies.

Conclusions:

  • The described method provides a valuable tool for investigating transcription regulation in specific neuronal populations.
  • This advancement opens new avenues for exploring cell type identity and function in the brain at a molecular level.
  • The protocol facilitates the use of genome-wide assays in neurobiology, advancing our understanding of the mammalian brain.