核酸酶靶向切割和释放 (CUT&RUN)是建立在免疫探测技术上的染色质图谱分析方法。在CUT&RUN中，融合蛋白AG-微球菌核酸酶（pAG-MNase），选择性原位切割抗体结合的染色质（Figure 1）。该方法与二代测序（NGS）兼容，可提供组蛋白翻译后修饰（PTMs）和染色质相关蛋白（如转录因子[TFs]）的高分辨率全基因组图谱。
FIGURE 1 Overview of the CUTANA™ CUT&RUN protocol.
产品优势
CUTANATM Library Prep Kit 是第一个专门为CUT&RUN分析开发的文库构建试剂盒。
•对CUT&RUN的方法进行了优化，对比多用途或ChIP-seq文库构建试剂盒更具优势。
•对于CUT&RUN产生的有限输入，工作流程非常稳定，为使用0.5-10 ng DNA的Illumina® NGS提供了高质量的文库。
•试剂盒包含CUT&RUN文库制备所需的所有材料（酶、引物、DNA纯化磁珠、缓冲液和PCR管）。
•可轻松搭配CUTANA™ CUT&RUN试剂盒（EpiCypher 14-1048）或CUT&RUN Protocol （EpiCypher.com/protocols）使用，实现工作流程集成，高通量检测和保证结果的可靠性，降低实验成本。
•可以有效制备组蛋白PTMs和染色质相关蛋白（如TFs）CUT&RUN DNA的文库。
Multiplexing Primers
Primer Set 1 includes i5 primers 1-8 and i7 primers 1-6
Primer Set 1 includes i5 primers 1-8 and i7 primers 7-12
保存条件
OPEN KIT IMMEDIATELY and store components at room temperature and -20°C as indicated (see Kit Manual for full instructions). Stable for 6 months upon date of receipt.
Room Temperature (RT) | -20℃ |
8-strip Tubes | End Prep Enzyme | Ligation Enhancer |
SPRIselectreagent manufactured by Beckman Coulter, Inc. | End Prep Buffer | High Fidelity 2X PCR Master Mix |
0.1X TE Buffer | Adapter for Illumina® | U-Excision Enzyme |
| Ligation Mix | i5 and i7 Primers |
数据示例
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FIGURE 1： CUT&RUN DNA Fragment Size Distribution Analysis. CUT&RUN was performed as described above. Library DNA was analyzed by Agilent TapeStation® , which confirmed that mononucleosomes were predominantly enriched in CUT&RUN (~300 bp peaks represent 150 bp nucleosomes + sequencing adapters). Peaks at ~380 bp correspond to the SNAP-CUTANA™ K-MetStat Panel of spike-in controls(EpiCypher 19-1002). |
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FIGURE 2： Representative Gene Browser Tracks. CUT&RUN was performed as described above. A representative 674 kb window at the SEPTIN5 gene is shown for three replicates (”Rep”) of IgG and H3K4me3 antibodies, as well as individual tracks for H3K27me3 and the transcription factor CTCF, demonstrating the robustness and reproducibility of the workflow with a variety of targets. Sequencing libraries prepared with the CUTANA CUT&RUN Library Prep kit produced the expected genomic distribution for each target. Images were generated using the Integrative Genomics Viewer (IGV, Broad Institute) |
订购详情
货号 | 产品名称 | 规格 |
14-1001 | CUTANA™ CUT&RUN Library Prep Kit with Primer Set 1 | 48 Reactions |
14-1002 | CUTANA™ CUT&RUN Library Prep Kit with Primer Set 2 | 48 Reactions |
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