Revision 8
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

C&T

:

:

:

:

Product Information

Storage

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

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

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.

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.

  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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

C&T: CUT&Tag

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 8
#47415

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

CUT and Tag Image 1: CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems Expand Image
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.
CUT and Tag Image 2: CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems Expand Image
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.
CUT and Tag Image 3: CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems Expand Image
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.