Diagenode

iDeal CUT&Tag kit for Histones

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Catalog Number
Format
Price
C01070020
24 rxns
$1,540.00
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. The protocol is optimized for native cells (10,000-300,000 cells per reaction) and can be completed within 2.5 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 marks
    • 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

  • 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.

    Antibody package for CUT & Tag

  • Examples of results

    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.

  •  Documents
    Chromatin Brochure BROCHURE
    Whether you are experienced or new to the field of chromatin immunoprecipitation, Diagenode has e...
    Download
    iDeal CUT&Tag kit for Histones MANUAL
    Download
  •  Publications

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