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Improved Protocol for Chromatin Immunoprecipitation from Mouse Skeletal Muscle
Published on: November 6, 2017
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Chromatin Immunoprecipitation in Skeletal Myoblasts
Vinay Kumar Rao1,2, Shilpa Rani Shankar1, Reshma Taneja3
1Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
Methods in Molecular Biology (Clifton, N.J.)
|October 28, 2018
Summary
This study details a chromatin immunoprecipitation (ChIP) protocol for analyzing DNA-protein interactions in skeletal myoblasts. It optimizes steps for identifying histone modifications and transcription factor binding genome-wide.
Area of Science:
- Molecular Biology
- Genomics
- Epigenetics
Background:
- Chromatin immunoprecipitation (ChIP) is a key technique for investigating DNA-protein interactions.
- ChIP facilitates unbiased, genome-wide analysis of transcriptional regulation in biological processes like cellular differentiation.
Purpose of the Study:
- To provide a detailed, step-by-step protocol for ChIP in skeletal myoblasts.
- To identify histone modifications, transcription factor binding, and co-factor interactions with chromatin.
- To offer guidance on critical experimental steps and common pitfalls.
Main Methods:
- Development and optimization of a chromatin immunoprecipitation (ChIP) protocol.
- Specific focus on cell harvesting, chromatin sonication, and immunoprecipitation procedures.
- Application in skeletal myoblasts for studying DNA-protein interactions.
Main Results:
- A refined ChIP protocol for skeletal myoblasts.
- Identification of key steps and potential pitfalls in the ChIP process.
- Enables genome-wide analysis of epigenetic modifications and transcription factor binding.
Conclusions:
- The described ChIP protocol is effective for studying DNA-protein interactions in skeletal myoblasts.
- This method allows for genome-wide assessment of transcriptional changes during differentiation.
- The protocol addresses critical steps to ensure successful and reproducible results.
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