Measuring B Cell Chromatin Accessibility by One-Step ATAC-seq

Methods Mol Biol. 2024:2826:55-63. doi: 10.1007/978-1-0716-3950-4_5.

Abstract

The Assay for Transposase Accessible Chromatin (ATAC)-seq protocol is optimized to generate global maps of accessible chromatin using limited cell inputs. The Tn5 transposase tagmentation reaction simultaneously fragments and tags the accessible DNA with Illumina Nextera sequencing adapters. Fragmented and adapter tagged DNA is then purified and PCR amplified with dual indexing primers to generate a size-specific sequencing library. The One-Step workflow below outlines the Tn5 nuclei transposition from a range of cell inputs followed by PCR amplification to generate a sequencing library.

Keywords: ATAC-seq; Chromatin accessibility; Epigenetic; Gene regulation; Sequencing library; Tn5 transposase.

MeSH terms

  • Animals
  • B-Lymphocytes* / metabolism
  • Chromatin Immunoprecipitation Sequencing / methods
  • Chromatin* / genetics
  • Chromatin* / metabolism
  • DNA / genetics
  • Gene Library
  • High-Throughput Nucleotide Sequencing* / methods
  • Humans
  • Polymerase Chain Reaction / methods
  • Sequence Analysis, DNA / methods
  • Transposases* / genetics
  • Transposases* / metabolism

Substances

  • Chromatin
  • Transposases
  • Tn5 transposase
  • DNA