CUT&Tag技术的核心是在Tn5上融合了protein A/G抗体结合功能域的转座体pAG-Tn5。在进行CUT&Tag实验时，首先进行靶蛋白特异性抗体（一抗）孵育，使抗体进入细胞与靶蛋白结合。为了放大信号，同理接着进行二抗孵育。最后孵育pAG-Tn5转座体，使得转座体进入细胞并与抗体结合，这样就把转座体间接的固定在靶蛋白上，随后...
CUT&Tag是研究蛋白质与DNA相互作用的一种较新的分子生物学方法,与ChIP-Seq和CUT&RUN相比具有一些优势。 1 起始量低 在最早的CUT&Tag文献中,研究人员用了60个细胞来分析整个基因组中的H3K27me3基因谱。之所以CUT&Tag能用如此低的细胞数,是因为pA-Tn5可以...
CUT&Tag 技术是基于在单个细胞中进行染色质分析,但受到可以分析的细胞数量的限制。 文章中描述了一种用于 CUT&Tag 的组合条形码策略(a combinatorial barcoding strategy),该策略利用纳米孔分配器 (nanowell dispenser)进行简单,高分辨率,高通量的单细胞染色质分析。 文章给出了比较详细的实验 protocol sciCUT&Tag,轻...
Ye Zheng, Kami Ahmad, Steven Henikoff. CUT&Tag Data Processing and Analysis Tutorial. Protocol .io, 2020. Kaya-Okur HS, Wu SJ, Codomo CA, et al. CUT&Tag for efficient epigenomic profiling of small samples and single cells. Nat Commun. 2019;10(1):1930....
CUT&Tag-IT™ Assay Kit商品 应用方向 1.组蛋白修饰/DNA 结合蛋白分析 使用CUT&Tag 能够达到高效、低成本的组蛋白修饰分析,并且适用于不同类型的靶蛋白和转录因子。 Kaya-Okur H S , Janssens D H , Henikoff J G , et al. Efficient low-cost chromatin profiling with CUT&Tag[J]. Nature Protocol,...
PCR tubes or plate and PCR Machine CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems Protocol SAFE STOP This is a safe stopping point in the protocol, if stopping is necessary. I. Low Plexity Pooling Guidelines: The dual index primer strategy utilizes two 8 base indices...
CUT&Tag细胞起始量通常为106,诺禾致源CUT&Tag可以超低量——1000个细胞建库。 一抗:H3K4me2 细胞量:105、104、103 结果: (1)reads在TSS附近富集; (2)1000细胞起始量检出peak数目与104、105细胞数的peak检出数目相当; (3)皮尔逊相关系数大于0.95,表明不同起始量...
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CUT&Tag is based on the same principles as ChIP-Seq, but with several changes to the protocol that are advantageous in certain situations. Instead of the sonication of fixed chromatin and immunoprecipitation steps performed in ChIP-Seq protocols, in CUT&Tag, fresh (not frozen) unfixed cells are...
This protocol paper aims to provide the new researchers with the full details of using Cleavage Under Targets and Tagmentation (CUT&Tag) to profile the genomic locations of chromatin binding factors, histone marks, and histone variants. CUT&Tag protocols function very well with mouse myoblasts and...