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Low-Input MNase Accessibility of Chromatin (Low-Input MACC)
Mattia Lion1,2, Michael Y Tolstorukov3, Marjorie A Oettinger1,2
1Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts.
Current Protocols in Molecular Biology
|June 26, 2019
Summary
A new low-input MNase accessibility (MACC) technique enables genome-wide chromatin profiling from minimal cell samples. This robust method enhances the study of chromatin structure and nucleosome occupancy in various research and clinical settings.
Area of Science:
- Molecular Biology
- Genomics
- Epigenetics
Background:
- Understanding dynamic chromatin structure is crucial for molecular biology.
- Existing techniques for chromatin profiling and nucleosome occupancy assessment are limited.
- Simultaneous measurement of chromatin opening, compaction, and nucleosome occupancy is needed.
Purpose of the Study:
- To present a low-input MNase accessibility (MACC) procedure.
- To extend the utility of the original MACC assay for genome-wide chromatin analysis.
- To enable high-quality chromatin profiling from very low cell numbers.
Main Methods:
- Development of a low-input MNase accessibility (MACC) protocol.
- Utilizing minimal cell input (as few as 50 cells).
- Genome-wide profiling of chromatin opening, compaction, and nucleosome occupancy.
Main Results:
- High-quality data generation from extremely low cell numbers.
- Successful simultaneous measurement of chromatin accessibility and nucleosome occupancy.
- Significant improvements in speed and robustness compared to the original MACC method.
Conclusions:
- Low-input MACC significantly expands the applicability of MACC assays.
- This method is ideal for limited cell samples, such as those from FACS or clinical settings.
- The enhanced MACC procedure offers a more rapid and robust approach to chromatin analysis.
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