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

DOI:
10.3791/56013-v

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10:04 min

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July 06, 2017

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, ,
1Department of Biochemistry and Molecular Biology,University of Texas Health Science Center at Houston

Chapters

  • 00:05Title
  • 01:04Nuclei Isolation from Cross-linked Skeletal Muscle
  • 04:11Sonication
  • 05:14ChIP
  • 07:53Results: Purification of High-quality Nuclei from Cross-linked Skeletal Muscle Suitable for Chromatin Immunoprecipitation
  • 08:51Conclusion

Summary

Automatic Translation

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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.

Tags

Cross-linked Skeletal MuscleChromatin ImmunoprecipitationNuclei IsolationFiltration-based MethodMuscle Genomic StudiesTime-sensitive ContextFormaldehyde Cross-linkingDounce HomogenizerProtease Inhibitors

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