供应商 :近岸蛋白
规格 :12次/24次
正值近岸蛋白上市一周年之际,为感恩广大客户的支持,即日起至2023年10月31日,近岸蛋白精选靶点及因子类蛋白、分子生物学试剂限时优惠大促销,多买多送,等你参加!
目录号:N259-YH01
产品描述:
CUT&Tag技术介绍
Cleavage Under Target & Tagmentation (CUT&Tag) 是一种新兴的 DNA- 蛋白质互作研究方法,可以用于替代ChIP实验。CUT&Tag以融合了protein A/G的转座酶为核心,融合蛋白通过protein A/G与抗体结合,使得转座酶被栓系在靶位点周围,从而在目的位点附近进行酶切反应,并同时引入二代测序所必需的接头序列,产物DNA经过后续提取与PCR扩增后,得到直接用于测序的文库。CUT&Tag 与传统的 ChIP-seq技术相比较,实验不需要超声打断,也不需要传统的连接法添加测序接头,具有操作简单、周期短、信噪比高、重复性好、细胞用量少等优势。
图1 ChIP 与CUT&Tag 的比较
存在问题
自2019年CUT&Tag技术诞生以来,经过大量的实践,发现该技术在组蛋白修饰及部分转录因子与DNA互作研究中可以获得高信噪比的结果,但是针对转录因子及辅因子等低丰度位点,较难得到理想的实验结果。
近岸蛋白创新开发高浓度甲醛交联方案
针对此问题,近岸蛋白开发了可对细胞进行高浓度甲醛交联的CUT&Tag实验流程。在该流程中,与传统ChIP-seq一样,首先对细胞进行高浓度甲醛交联处理,再经过细胞透化后,即可进入常规CUT&Tag实验流程。该流程兼顾高浓度甲醛交联固定的优势及CUT&Tag实验细胞用量少、操作简单、信噪比高的优点,为转录因子-DNA互作研究提供一种新的思路,这种需要甲醛交联的 CUT&Tag 实验被称为fcCUT&Tag(formaldehyde crosslinking CUT&Tag)。
NovoNGS® CUT&Tag® 4.0 High-Sensitivity Kit (for Illumina®)试剂盒包含常规非交联CUT&Tag 及甲醛交联 fcCUT&Tag 实验中所需的各种缓冲液、ConA磁珠、DNA纯化磁珠、引物等全套建库试剂,可以用于组蛋白修饰和转录因子的染色质结合状态研究。与传统的CUT&Tag试剂盒相比,该试剂盒既可以对自然状态的细胞进行蛋白质 -DNA 互作研究,也可以对高浓度甲醛交联的细胞进行蛋白质 -DNA 互作研究,一盒两用。
产品用途:
DNA-蛋白质互作研究。在常见的哺乳动物细胞上用于替代传统的ChIP-Seq 技术,特别适用于细胞数量较少的样本,也可应用于经过处理的植物和酵母样本上。
基本原理:细胞被固定在刀豆蛋白磁珠(ConA磁珠)表面,然后加入洋地黄皂苷(digitonin)透化细胞膜,依次进行一抗、二抗、转座体的孵育,随后加入Mg2+激活转座酶进行片段化反应,反应产物经过DNA提取、PCR富集完成二代测序文库的构建。对于人源细胞常规的组蛋白修饰,仅需进行3-5 M reads的测序量就能得到高分辨率的全基因组图谱。
产品性能和应用实例:
FcCUT&Tag与CUT&Tag实验数据对比:
结果表明:对于转录因子而言,利用高浓度甲醛交联处理后,实验获得的peak数更多,背景更低,信噪比更高。
FOR RESEARCH USE ONLY
该产品被引用文献
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