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Nucleosome Positioning - Usage Guide

Overview

Extract nucleosome positions from ATAC-seq fragment size patterns using ATACseqQC or NucleoATAC to understand chromatin structure at promoters and regulatory regions.

Prerequisites

BiocManager::install(c('ATACseqQC', 'TxDb.Hsapiens.UCSC.hg38.knownGene'))
pip install nucleoatac

Quick Start

Tell your AI agent what you want to do:

  • "Analyze nucleosome positioning from my ATAC-seq data"
  • "Generate fragment size distribution to check nucleosome periodicity"

Example Prompts

Fragment Size Analysis

"Plot the fragment size distribution from my ATAC-seq BAM to visualize nucleosome patterns"

Nucleosome Calling

"Call nucleosome positions from my ATAC-seq data using NucleoATAC"

TSS Nucleosome Patterns

"Analyze nucleosome positioning around TSS to visualize NFR and +1/-1 nucleosomes"

NFR Analysis

"Extract nucleosome-free region reads and analyze coverage at promoters"

What the Agent Will Do

  1. Calculate fragment size distribution to verify nucleosome periodicity
  2. Separate reads by fragment size (NFR, mono-, di-, tri-nucleosomal)
  3. Call nucleosome positions using NucleoATAC or ATACseqQC
  4. Generate nucleosome occupancy signal tracks
  5. Visualize nucleosome patterns around TSS

Fragment Size Interpretation

Fragment Size Origin Meaning
<100 bp Nucleosome-free Active promoter/enhancer
180-247 bp Mono-nucleosome Well-positioned nucleosome
315-473 bp Di-nucleosome Regular spacing
558-615 bp Tri-nucleosome Chromatin structure

Nucleosome Pattern Interpretation

Pattern Meaning
Strong NFR peak Active promoter/enhancer
Regular spacing Well-positioned nucleosomes
Fuzzy positioning Dynamic chromatin

Tips

  • Paired-end data is required for accurate fragment sizing
  • Use properly deduplicated BAM files
  • Filter for high-quality fragments (MAPQ > 30)
  • High mitochondrial contamination can skew fragment distributions
  • NucleoATAC provides base-pair resolution nucleosome positions
  • ATACseqQC is useful for V-plots around TSS showing nucleosome phasing