CUT&Tag for Efficient Epigenomic Profiling of Frozen Tissues.

Area of Science:

• Molecular Biology
• Genomics
• Epigenetics

Background:

• Profiling chromatin components is crucial for understanding gene regulation.
• Existing methods may have limitations in resolution or sample compatibility.
• Cleavage Under Targets and Tagmentation (CUT&Tag) offers a high-resolution approach.

Purpose of the Study:

• To detail a robust protocol for generating CUT&Tag sequencing libraries.
• To demonstrate the applicability of the protocol across diverse fresh and frozen tissue types.
• To enable high-resolution profiling of chromatin components.

Main Methods:

• Nuclei isolation from fresh or frozen tissues.
• Antibody binding and pA-Tn5 adapter complex incubation.
• Tagmentation, DNA extraction, PCR amplification, and library purification.

Main Results:

• A comprehensive nine-step CUT&Tag workflow is described.
• The protocol successfully generated sequencing libraries from various tissue types.
• High-resolution profiling of chromatin components was achieved.

Conclusions:

• The presented CUT&Tag protocol is versatile and effective for fresh and frozen tissues.
• This method provides high-resolution insights into chromatin organization.
• The protocol facilitates the study of diverse chromatin components across different tissue sources.