Protocol for genomic mapping of chromatin targets using high-throughput CUT&RUN

Area of Science:

• Molecular Biology
• Genomics
• Epigenetics

Background:

• Understanding protein distribution in chromatin is crucial for gene regulation, cellular identity, and disease.
• Transcription factors, chromatin modifiers, and histone modifications are key regulators of chromatin state.

Purpose of the Study:

• To present a high-throughput, 96-well plate-based cleavage under targets and release using nuclease (CUT&RUN) protocol.
• To enable high-resolution genomic mapping of proteins and epigenomic factors.

Main Methods:

• High-throughput CUT&RUN protocol in a 96-well plate format.
• Detailed steps for targeted DNA enrichment, library preparation, sequencing, and data analysis.

Main Results:

• Successful implementation of a scalable CUT&RUN protocol.
• High-resolution genomic mapping of proteins and epigenomic factors is achievable.

Conclusions:

• The developed protocol facilitates efficient and high-resolution mapping of chromatin-associated proteins.
• This method aids in understanding gene regulation, cellular identity, and disease development through epigenomic analysis.