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Related Experiment Video

Updated: Mar 12, 2026

Generation of Native Chromatin Immunoprecipitation Sequencing Libraries for Nucleosome Density Analysis
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Methods for Investigating DNA Accessibility with Single Nucleosomes.

M D Gibson1, M Brehove1, Y Luo1

  • 1The Ohio State University, Columbus, OH, United States.

Methods in Enzymology
|October 30, 2016
PubMed
Summary
This summary is machine-generated.

This study details single-molecule total internal reflection fluorescence (smTIRF) microscopy methods to observe how proteins access DNA within nucleosomes. These techniques are crucial for understanding DNA processing and protein-DNA interactions.

Keywords:
Fluorophore-labeled nucleosomesNucleosome accessibilitySingle-molecule fluorescenceTranscription factor-binding dynamics

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biophysics

Background:

  • Nucleosomes are the core structural units of eukaryotic DNA.
  • Understanding protein access to nucleosomal DNA is vital for DNA processing (transcription, replication, repair).

Purpose of the Study:

  • To describe methods for single-molecule total internal reflection fluorescence (smTIRF) microscopy.
  • To investigate protein binding dynamics within nucleosomes using smTIRF.

Main Methods:

  • Preparation of fluorophore-labeled nucleosomes.
  • Utilizing a single-molecule total internal reflection fluorescence (smTIRF) microscopy system.
  • Data acquisition and analysis protocols for smTIRF experiments.

Main Results:

  • Established protocols for smTIRF experiments on nucleosomes.
  • Demonstrated applicability for studying transcription factor binding.
  • Validated methods for observing protein-DNA interactions at the single-molecule level.

Conclusions:

  • smTIRF microscopy provides key insights into protein-DNA interactions within nucleosomes.
  • The described methods are versatile for studying various DNA-binding proteins.
  • This work facilitates research into fundamental DNA processes.