Diagenode

iDeal CUT&Tag kit for Histones
Compatible with histones and some non-histone proteins

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Catalog Number
Format
Price
C01070025
8 rxns
$560.00
Other format


CUT&Tag-sequencing (Cleavage Under Targets and Tagmentation) is a new alternative method to ChIP-seq combining antibody-targeted controlled cleavage by a protein A-Tn5 fusion with massively parallel DNA sequencing to identify the binding sites of DNA-associated proteins.

The Diagenode’s iDeal CUT&Tag kit for Histones provides an optimized protocol for a rapid chromatin profiling on histone marks and some non-histone proteins. The protocol is optimized for native cells (10,000-300,000 cells per reaction) and can be completed within 1.5 days. The kit includes all reagents for cell processing, including CoA beads, pA-Tn5 and the DNA purification module. The antibodies (secondary antibodies, control antibodies) as well as primer indexes for multiplexing must be purchased separately.

For a complete CUT&Tag protocol the following items must be purchased:

  • iDeal CUT&Tag kit for Histones – including all reagents for CUT&Tag workflow (buffers, pA-Tn5, CoA beads, DNA purification)
  • Antibody package for CUT&Tag: anti-rabbit or anti-mouse - including the secondary antibody, positive and negative control antibodies and primers
  • Primer indexes for tagmented libraries – for multiplexing up to 72 samples

iDeal CUT&Tag Kit features:

  • Rapid and easy chromatin profiling assay for histones and some non-histone proteins
    • No chromatin preparation
    • Easy sample handling due to ConA magnetic beads
    • Integrated library prep
  • Low cell number: 10,000-300,000 cells
  • Accurate amplification due to intermediate quantification step
  • High resolution and sensitivity
  • Lower sequencing depth

The quality of antibody used in CUT&Tag is one of the crucial factors for assay success. The antibodies with confirmed high specificity will target only the protein of interest, enabling real results. Check out our selection of antibodies validated in CUT&Tag.

Looking for a standalone pA-Tn5? Read more.

  • Method overview

    The iDeal CUT&Tag protocol involves the binding of cells on a solid phase ConA magnetic beads, allowing magnetic handling of the cells for the major steps of the protocol. Bead-bound cells are permeabilized, incubated with primary antibody against a target of interest and secondary antibody. Then, Diagenode’s protein pA-Tn5 Transposase - loaded is bound to the complex. Protein A guides Tn5 transposase on chromatin to the antibody attached to its target. Tn5 transposase is activated by Mg+2 ions to insert the sequencing adaptors into genomic regions of interest. DNA is then purified and the tagmented genomic regions of interest are amplified by PCR using Diagenode’s Primer Indexes for tagmented libraries.

      pA-Tn5 Antibody package for CUT & Tag

  • Examples of results - histone marks

    Successful CUT&Tag results showing a low background with high region-specific enrichment are presented below. Chromatin profiling has been performed on 50,000 K562 cells, using the Diagenode’s iDeal CUT&Tag kit for Histones (Cat. No. C01070020), the Antibody Package for CUT&Tag anti rabbit (Cat. No. C01070022), the 24 UDI for Tagmented Libraries (Cat.No. C01011034) and H3K4me3 (Cat. No. C15410003), H3K27me3 (Cat. No. C15410069) or H3H9me3 antibodies (Cat. No., C15410193) as indicated. The libraries were sequenced on Illumina’s NovaSeq6000 in 2x50 bp mode.


    CUT&Tag-sequencing CUT&Tag kit for Histones Cleavage Under Targets and Tagmentation

    Figure 1. Typical library profiles (Agilent Fragment traces) generated by the iDeal CUT&Tag protocol using 50,000 K562 cells and H3K27me3 (top), H3K4me4 (middle) primary antibodies and IgG control (bottom). Sharp peak at around 40 bp is an excess of free oligonucleotide used for pA-Tn5 loading.



    iDeal CUT&Tag kit for Histones

    Figure 2. Enrichments at TSS of the CUT&Tag libraries. The heatmap shows the enrichment around 3kb upstream and downstream of the TSS for H3K4me3. H3K4me3 as an active chromatin mark is associated with active promoters shows a narrow enrichment pattern.




    iDeal CUT&Tag experiments of K562 cells

    Figure 3. Sequencing profiles of the CUT&Tag libraries. Integrative genomics viewer (IGV) visualization of CUT&Tag experiments using 50,000 of K562 cells and H3K4me3 (blue), H3K27me3 (red) or H3K9me3 (green).




    CUT&Tag experiments using 10,000 K562 cells

    Figure 4. Sequencing profiles of the CUT&Tag libraries. Integrative genomics viewer (IGV) visualization of CUT&Tag experiments using 50,000 (blue) or 10,000 (red)) of K562 cells and H3K27me3 antibody.

  • Examples of results - transcription factors

    Diagenode's iDeal CUT&Tag Kit for Histonesdeveloped for an efficient chromatin profiling on histone marks, can be used for profiling of some transcription factors and co-factors using a mild fixation as described in the manual. 

    Successful CUT&Tag results using CTCF and Suz12 antibodies and iDeal CUT&Tag Kit for Histones. 

    Chromatin profiling of Suz12 has been performed on 50,000 of fixed Mouse ES-E14TG2a cells (kindly provided by Luciano Di Croce, CRG, Spain) accordingly to the protocol of Diagenode’s iDeal CUT&Tag kit for Histones (Cat. No. C01070020).   Antibody Package for CUT&Tag anti rabbit (Cat. No. C01070022) and the 24 UDI for Tagmented Libraries (Cat. No. C01011034) were used. The libraries were sized using Fragment Analyzer (Agilent) (triplicates, Figure 1, top). Relative enrichment has been confirmed by qPCR using know positive (T_Bra)  and negative (Actb)  loci  (Figure 1, bottom). Libraries were sequenced on Illumina’s NovaSeq6000 in 2x50 bp mode.


    Figure 1. Typical library profiles (top) and relative enrichment (bottom) generated by the iDeal CUT&Tag protocol using 50,000 ES-E14TG2a cells and Suz12 Antibody and IgG control (bottom). Sharp peak at around 40 bp is an excess of free oligonucleotide used for pA-Tn5 loading which does not interfere with the sequencing.

    Pictures of representative loci were taken on Integrative Genome Viewer (IGV) for CTCF demonstrating the presence of high signal at promoter regions.

    Pictures of representative loci were taken on Integrative Genome Viewer (IGV) for SUZ12 and H3K27me3 triplicate samples, showing an overlap between the signal from both proteins, as part of the Polycomb Repressive Complex.

    Figure 2. IGV snapshots

    Figure 3. Heatmap around the TSS
    Heatmap of the CTCF signal around the transcription start sites (TSS) of each gene present in the murine mm10 reference genome.

    Figure 4. CTCF motif
    Presence of specific transcription factor binding motifs in the regions identified as CTCF peaks. The top3 motifs are corresponding to the CTCF binding motif.

    Figure 5. CTCF motif density
    Location and density of the CTCF binding motif with respect to the center of the identified CTCF peaks.

  •  Documents
  •  Safety sheets
    iDeal CUT&Tag kit for Histones SDS US en Download
    iDeal CUT&Tag kit for Histones SDS GB en Download
    iDeal CUT&Tag kit for Histones SDS BE fr Download
    iDeal CUT&Tag kit for Histones SDS DE de Download
    iDeal CUT&Tag kit for Histones SDS BE nl Download
    iDeal CUT&Tag kit for Histones SDS JP ja Download
    iDeal CUT&Tag kit for Histones SDS FR fr Download
    iDeal CUT&Tag kit for Histones SDS ES es Download
  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: iDeal CUT&Tag kit for Histones
    Compatible with histones and some non-histone proteins (Diagenode Cat# C01070025)
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