CUT&Tag 技术可高效进行冰冻组织的表观基因组分析。

CUT&Tag for Efficient Epigenomic Profiling of Frozen Tissues.

机构信息

Institute of Systems Biomedicine, Department of Pathology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China.

State Key Laboratory of Oral Diseases & National Center for Stomatology & National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China.

出版信息

Methods Mol Biol. 2024;2846:181-189. doi: 10.1007/978-1-0716-4071-5_12.

DOI:10.1007/978-1-0716-4071-5_12

PMID:39141237

Abstract

Cleavage Under Targets and Tagmentation (CUT&Tag) provides high-resolution sequencing libraries for profiling diverse chromatin components. This protocol details the steps to generate CUT&Tag libraries from fresh or frozen tissues. This CUT&Tag workflow has nine main steps: isolation of nuclei from tissues, binding of nuclei to Concanavalin A-coated beads, binding of the primary antibody, binding of the secondary antibody, binding pA-Tn5 adapter complex, tagmentation, DNA extraction, PCR, and post-PCR cleanup and size selection. This protocol enabled us to generate and sequence CUT&Tag libraries across a broad range of fresh and frozen tissue types.

摘要

靶向切割和标签化（CUT&Tag）为分析多种染色质成分提供了高分辨率测序文库。本方案详细介绍了从新鲜或冷冻组织中生成 CUT&Tag 文库的步骤。该 CUT&Tag 工作流程主要包括九个步骤：组织细胞核的分离、细胞核与伴刀豆球蛋白 A 包被珠的结合、一抗的结合、二抗的结合、pA-Tn5 接头复合物的结合、标签化、DNA 提取、PCR 以及 PCR 后清洗和大小选择。该方案使我们能够在广泛的新鲜和冷冻组织类型中生成和测序 CUT&Tag 文库。