CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems

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产品信息

简单描述

Next generation sequencing (NG-seq) is a high throughput method that can be used downstream of the Cleavage Under Targets and Tagmentation (CUT&Tag) assay to identify and quantify target DNA enrichment across the entire genome. The CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems kit is ideally suited for multiplex sample preparation for NG-seq on the Illumina systems platform. This kit can be used to generate up to 96 distinct, barcoded CUT&Tag DNA libraries that can be combined into a single sequencing reaction. This product provides enough reagents to support up to 96 DNA sequencing libraries. This product is compatible with CUT&Tag DNA samples generated by CUT&Tag Assay Kit #77552 or CUT&Tag pAG-Tn5 (Loaded) #79561 and DNA samples from other tagmentation assays, such as ATAC-seq. This product is not compatible with SimpleChIP® Chromatin IP Kits (#9003, #9005, #56383) or the CUT&RUN Assay Kit #86652.

应用

目标/特异性

Specificity/Sensitivity

This kit can generate DNA libraries using CUT&Tag DNA. Note: While CUT&Tag DNA libraries generated for histone modifications typically show robust signal in Agilent Bioanalyzer or TapeStation systems analysis, libraries generated for non-histone proteins such as transcription factors and cofactors often have very weak or even no visible signal using Bioanalyzer or TapeStation systems, but still generate NG-sequencing results with high mapping rates, high numbers of identified binding peaks, and decent signal-to-noise ratios across the whole genome.

Species Reactivity:

All Species Expected

背景

Similar to Cleavage Under Targets and Release Using Nuclease (CUT&RUN), Cleavage Under Targets and Tagmentation (CUT&Tag) is a powerful technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1-3). CUT&Tag has many of the same advantages as the CUT&RUN assay in that it provides a rapid, robust, and true low cell number protocol for detection of protein-DNA interactions in the cell. In addition, the CUT&Tag assay adds an in situ adaptor DNA ligation step carried out by the pAG-Tn5 enzyme, in which an adaptor DNA is ligated directly to antibody-targeted chromatin DNA fragments in the cell. As a result, subsequent DNA library preparation is much faster and easier than library preparation following the CUT&RUN assay, free from DNA end repair, A-tailing, and adaptor ligation in vitro. CUT&Tag works very well for analyzing histone modifications, in addition to mapping some transcription factor and cofactor binding. 1.Kaya-Okur, H.S. et al. (2019) Nat Commun 10, 1930. 2.Kaya-Okur, H.S. et al. (2020) Nat Protoc 15, 3264-3283. 3.Henikoff, S. et al. (2021) Bio Protoc 11, e4043.

翻译后修饰

unmodified

制备和贮存

保存方式

Store all components at -20ºC. This product is stable for 18 months if stored properly.

参考图片

Figure 3. CUT&Tag was performed with HCT 116 cells and Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb #9751, using CUT&Tag Assay Kit #77552. The DNA library was prepared using indexes i504 and i703 from CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems. The figure shows binding across chromosome 12 (upper), including GAPDH (lower), a known target gene of H3K4me3.

Figure 1. CUT&Tag was performed with NCCIT cells and JARID2 (D6M9X) Rabbit mAb #13594, using CUT&Tag Assay Kit #77552. The DNA library was prepared using indexes i501 and i705 from CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems. The figure shows binding across HOXA (upper) and HOXD (lower), known target genes of JARID2.

Figure 2. CUT&Tag was performed with HCT 116 cells and TCF4/TCF7L2 (C48H11) Rabbit mAb #2569, using CUT&Tag Assay Kit #77552. The DNA library was prepared using indexes i502 and i706 from CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems. The figure shows binding across chromosome 8 (upper), including MYC (lower), a known target gene of TCF4.

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货号：

47415S

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