周年庆感恩买三赠一(即日起至2022.12.31)
目录号:N259-YH01-01B
产品描述:
Cleavage Under Target & Tagmentation (CUT&Tag)是一种新兴的DNA-蛋白质互作研究方法。CUT&Tag以融合了protein A/G的Tn5转座酶为核心,融合蛋白通过protein A/G与抗体结合,使得Tn5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,操作简单、周期短、信噪比高、重复性好、细胞用量少,为极低起始细胞量和单细胞测序研究提供了可能性。
NovoNGS® CUT&Tag 3.0 High-Sensitivity Kit (for Illumina®) 试剂盒包含CUT&Tag实验中孵育抗体、转座体、构建文库的相关试剂,可以用于组蛋白修饰和转录因子的染色质结合状态研究。本试剂盒包含高活性的ChiTag® pAG-Tn5,相较于最初版本的pA-Tn5转座酶,pAG结构域对兔、鼠等多种来源抗体具有广谱的强亲和性,在实验抗体的选择上具有更大的空间。
针对CUT&Tag打断产物DNA提取过程中小片段丢失的问题,本试剂盒提供优化的Tagment DNA Extract Beads,相较于常规的片段分选磁珠可以完整的回收CUT&Tag产物中的小片段DNA,大大简化了酚氯仿-乙醇沉淀法的繁琐操作。此外,本试剂盒还提供了阳性抗体H3K4me3(兔抗)以及相应的二抗(羊抗兔IgG),用于阳性对照实验(仅B包装提供阳性抗体)。
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3-5M 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:
■ ChiTag® pAG-Tn5的酶切活性:
■ 不同细胞数量样本上的CUT&Tag结果:
■ 不同抗体的CUT&Tag结果:
■ 不同细胞系的CUT&Tag结果:
转座体单链接头序列:
Adaptor1: 5’-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-3’
Adaptor2: 5’-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG -3’
ME: 5’-pCTGTCTCTTATACACATCT-NH2-3’
转座体单链接头序列:
5’-AATGATACGGCGACCACCGAGATCTACAC-Index2(i5)-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-xxxxxxxxx-CTGTCTCTTATACACATCTCCGAGCCCACGAGAC-Index1(i7)-ATCTCGTATGCCGTCTTCTGCTTG-3’,其中-xxxxxxxxx-为插入序列,两端为文库结构。
产品引用文献:
1、CRISPR screening identifies CDK12 as a conservative vulnerability of prostate cancer[J]. Hanqi Lei et al. Cell Death and Disease. 2021.
2、Inhibition of Syk promotes chemical reprogramming of fibroblasts via metabolic rewiring and H2S production[J]. Weiyun Wang et al. Embo Journal. 2021.
3、SS18 regulates pluripotent-somatic transition through phase separation[J]. Junqi Kuang et al. Nature Communications. 2021.
4、N6-methyladenosine modification of MALAT1 promotes metastasis via reshaping nuclear speckles[J]. Xinyu Wang et al. Developmental Cell. 2021.
5、The chromatin accessibility landscape reveals distinct transcriptional regulation in the induction of human primordial germ cell-like cells from pluripotent stem cells[J]. Xiaoman Wang et al. Stem Cell Reports. 2021.
6、Cellular senescence in hepatocellular carcinoma induced by a long non-coding RNA-encoded peptide PINT87aa by blocking FOXM1-mediated PHB2[J]. Xiaohong Xiang et al. Theranostics. 2021.
7、Gastrin, via Activation of PPARα, Protects the Kidney Against Hypertensive Injury[J]. Daqian Gu et al. Clinical Science. 2021.
8、Alpha‐fetoprotein accelerates the progression of hepatocellular carcinoma by promoting Bcl‐2 gene expression through an RA‐RAR signalling pathway[J]. Chao Zhang et al. Journal of Cellular and Molecular Medicine. 2020.
9、Pharmacological preconditioning by TERT inhibitor BIBR1532 confers neuronal ischemic tolerance through TERT-mediated transcriptional reprogramming[J]. Xuemin Xie et al. Journal of Neurochemistry. 2021.
10、A novel lncRNA linc-AhRA negatively regulates innate antiviral response in murine microglia upon neurotropic herpesvirus infection.[J]. Yiliang Wang et al. Theranostics. 2021.
11、Aberrant Fluid Shear Stress Contributes to Articular Cartilage Pathogenesis via Epigenetic Regulation of ZBTB20 by H3K4me3.[J]. Yu Jin et al. Journal of Inflammation Research. 2021.
12、The Synovium Attenuates Cartilage Degeneration in KOA through Activation of the Smad2/3-Runx1 Cascade and Chondrogenesis-related miRNAs.[J]. Xiaoyi Zhao et al. Molecular Therapy: Nucleic Acids. 2020.
For research use only