CUT&Tag Using "Stress-Free" Con A-Conjugated Dynabeads
CUT&Tag Using "Stress-Free" Con A-Conjugated Dynabeads
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DOI：
10.1007/978-1-0716-2433-3_16
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发表时间：
2023
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影响因子：
--
通讯作者：
Y
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文献类型：
--
作者：
Fujiwara;Y ;Okada;Y
表观基因组研究已经采用各种方法来鉴定感兴趣的蛋白质的基因组位置，例如转录因子和组蛋白修饰。CUT&Tag是最近建立的用于表观基因组研究的方法，用于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。在CUT&Tag方法中，细胞被结合并保持在伴刀豆球蛋白A（con A）包被的磁珠上，然后蛋白A Tn 5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。在这里，我们描述了更新的切割和标签程序使用“自制”的con A-共轭磁珠。该方法没有不良的悬浮性和严重的聚集，因此提供了改进的灵敏度。
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.