CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415
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Product Information
Storage
Store all components at -20ºC. This product is stable for 18 months if stored properly.
Protocol
Product Description
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.
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
Background
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.
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For Research Use Only. Not For Use In Diagnostic Procedures.
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