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使用“无压力”伴刀豆球蛋白A偶联磁珠的CUT&Tag技术

CUT&Tag Using "Stress-Free" Con A-Conjugated Dynabeads.

作者信息

Fujiwara Yasuhiro, Okada Yuki

机构信息

Institute for Quantitative Biosciences, The University of Tokyo, Tokyo, Japan.

出版信息

Methods Mol Biol. 2023;2519:141-153. doi: 10.1007/978-1-0716-2433-3_16.

Abstract

Epigenome research has employed various methods to identify the genomic location of proteins of interest, such as transcription factors and histone modifications. CUT&Tag is a recently established method used in epigenome research to determine the genomic location of proteins of interest, such as transcription factors and histone modifications. In CUT&Tag method, cells are bound and hold on concanavalin A (con A)-coated magnetic beads, then a Protein-A Tn5 transposase fusion protein cuts the genome and inserts adapter sequences nearby the target protein. Here we describe the updated CUT&Tag procedure using "home-made" con A-conjugated magnetic beads. This method is free of poor suspendability and severe aggregation, hence providing improved sensitivity.

摘要

表观基因组研究采用了多种方法来确定感兴趣的蛋白质(如转录因子和组蛋白修饰)的基因组位置。CUT&Tag是表观基因组研究中最近建立的一种用于确定感兴趣蛋白质(如转录因子和组蛋白修饰)基因组位置的方法。在CUT&Tag方法中,细胞与伴刀豆球蛋白A(Con A)包被的磁珠结合并固定,然后一种蛋白A-Tn5转座酶融合蛋白切割基因组并在靶蛋白附近插入接头序列。在此,我们描述了使用“自制”Con A偶联磁珠的更新后的CUT&Tag程序。该方法没有悬浮性差和严重聚集的问题,因此提高了灵敏度。

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