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Genetics

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

4.7K Views

08:40 min

August 14th, 2020

August 14th, 2020

4,675 Views

0:05

Introduction

0:21

Antibody Incubation with Beads

1:35

Chromatin Immunoprecipitation (ChIP)

2:56

Enzymatic Reactions on Beads

4:59

Elution and Purification

5:43

Library Preparation

6:50

Results: ChIP-exo Produces High Mapping Resolution

8:04

Conclusion

Transcript

The main advantage of our ChIP-exo method is the improved mapping resolution of protein DNA interactions in the cell compared to the conventional chromatin immunoprecipitation method. To prepare protein G magnetic beads for ChIP, mix the magnetic

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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.

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