Journal
/
Genetics
/
High-Resolution Mapping of Protein-DNA Interactions in Mouse Stem Cell-Derived Neurons using Chromatin Immunoprecipitation-Exonuclease (ChIP-Exo)
JoVE Journal
Genetics
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Genetics
High-Resolution Mapping of Protein-DNA Interactions in Mouse Stem Cell-Derived Neurons using Chromatin Immunoprecipitation-Exonuclease (ChIP-Exo)

High-Resolution Mapping of Protein-DNA Interactions in Mouse Stem Cell-Derived Neurons using Chromatin Immunoprecipitation-Exonuclease (ChIP-Exo)

DOI:
10.3791/61124-v

•

08:40 min

•

August 14, 2020

•

,
1Department of Cell and Systems Biology,University of Toronto, 2Department of Biology,University of Toronto

Chapters

  • 00:05Introduction
  • 00:21Antibody Incubation with Beads
  • 01:35Chromatin Immunoprecipitation (ChIP)
  • 02:56Enzymatic Reactions on Beads
  • 04:59Elution and Purification
  • 05:43Library Preparation
  • 06:50Results: ChIP-exo Produces High Mapping Resolution
  • 08:04Conclusion

Summary

Automatic Translation

Automatic Translation

Precise determination of protein-binding locations across the genome is important for understanding gene regulation. Here we describe a genomic mapping method that treats chromatin-immunoprecipitated DNA with exonuclease digestion (ChIP-exo) followed by high-throughput sequencing. This method detects protein-DNA interactions with near base-pair mapping resolution and high signal-to-noise ratio in mammalian neurons.

Tags

Keywords: ChIP-exoProtein-DNA InteractionsChromatin ImmunoprecipitationHigh-resolution MappingMouse Stem Cell-derived NeuronsMagnetic BeadsBlocking SolutionIslet-1 AntibodySonicated LysatesWash BufferEnd Prep Reaction MixEnd Prep Enzyme Mix

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/61124?article_type=v

Mapping RNA-RNA Interactions Globally Using Biotinylated Psoralen
check_url/61124?article_type=v

Genome-wide Mapping of Protein-DNA Interactions with ChEC-seq in Saccharomyces cerevisiae
check_url/61124?article_type=v

Chromatin Immunoprecipitation (ChIP) in Mouse T-cell Lines
check_url/61124?article_type=v

Retroviral Scanning: Mapping MLV Integration Sites to Define Cell-specific Regulatory Regions
check_url/61124?article_type=v

High Resolution Fluorescent In Situ Hybridization in Drosophila Embryos and Tissues Using Tyramide Signal Amplification
check_url/61124?article_type=v

Mapping Genome-wide Accessible Chromatin in Primary Human T Lymphocytes by ATAC-Seq
check_url/61124?article_type=v

Promoter Capture Hi-C: High-resolution, Genome-wide Profiling of Promoter Interactions
check_url/61124?article_type=v

CRISPR-Mediated Reorganization of Chromatin Loop Structure
check_url/61124?article_type=v

High-Resolution Comparison of Bacterial Conjugation Frequencies
check_url/61124?article_type=v

Chromatin Immunoprecipitation Assay Using Micrococcal Nucleases in Mammalian Cells

Read Article