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Biochemistry

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JoVE Journal Biochemistry
A Filtration-based Method of Preparing High-quality Nuclei from Cross-linked Skeletal Muscle for Chromatin Immunoprecipitation
 

A Filtration-based Method of Preparing High-quality Nuclei from Cross-linked Skeletal Muscle for Chromatin Immunoprecipitation

Article DOI: 10.3791/56013-v • 10:04 min • July 6th, 2017
July 6th, 2017 •
1, 1, 1
1Department of Biochemistry and Molecular Biology, University of Texas Health Science Center at Houston

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We present a filtration-based protocol to isolate high-quality nuclei from cross-linked mouse skeletal muscle wherein we removed the need for ultracentrifugation, making it easily applicable. We show that chromatin prepared from the nuclei is suitable for chromatin immunoprecipitation and likely chromatin immunoprecipitation sequencing studies.

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Filtration-based Method High-quality Nuclei Cross-linked Skeletal Muscle Chromatin Immunoprecipitation Genomic Studies Muscle Tissues Circadian Field Time-sensitive Context Minimal Myofibril Contamination Cardiac Muscles Smooth Muscles Hind Limb Skeletal Muscle C57 Black Six Male Mice Phosphate-buffered Saline Centrifuge Pellet Volume Formaldehyde
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