[Cell Signaling Technology] 효과적인 chromatin profiling을 위한 더 강력해진 기술, CUT&Tag을 소개합니다.

Cell Signaling Technology CUT&Tag

Cell Signaling Technology에서 엄격히 검증된 CUT&Tag kit 및 검증된 antibody 제품을 만나보세요!

What is CUT&Tag?

Cleavage Under Targets & Tagmentation (CUT&Tag)은 CUT&RUN과 마찬가지로 chromatin mapping을 위한 경제적이고 신속한 방법으로 1차 항체, 2차 항체 및 protein A-Tn5(pA-Tn5) transpose를 사용하여 차세대 염기서열 분석(NGS)을 위한 게놈 전체의 단백질-DNA 상호 작용을 식별해 낼 수 있습니다.

Key Features of CUT&Tag

  • Sequencing adapter와 함께 준비된 pA-Tn5의 tagmentation에 의해 DNA library 준비를 간소화
  • DNA library 분석까지 단 1-2일만에 진행 가능
  • 간편하고 편리한 프로토콜
  • 높은 민감도로 낮은 sequencing depth(~ 200만 reads)가 가능하여 분석 비용의 절감
  • 낮은 background signal로 강력한 데이터를 생성
  • Chromatin을 sonication 하지 않기 때문에 single cell 실험(scCUT&Tag)에 적용 가능

What Makes CUT&Tag Different

전통적인 ChIP와 ChIP-seq는 sonication 및 cross-linking된 chromatin을 사용하는 반면 CUT&Tag와 CUT&RUN은 native chromatin을 profiling하는 데 사용할 수 있습니다. 그러나 CUT&Tag은 CUT&RUN과 다르게 대부분의 library 준비가 in vivo에서 이루어지고 CUT&RUN과 동등한 수준의 데이터를 제공하며 single cell CUT&Tag를 가능하게 하므로 CUT&RUN보다 빠릅니다.

Cell Signaling Technology CUT&Tag

Cell Signaling Technology CUT&Tag Assay Kit #77552

  • 24번의 CUT&Tag assay을 진행할 수 있는 구성품(Concanavalin A bead, pAG-Tn5, buffer, 시약, Column, positive control, negative control, spike-in control, secondary antibody 등)이 포함
  • Next-generation sequencing (NG-seq)과 호환
  • 간편한 프로토콜 및 troubleshooting 제공
CUT&Tag Assay Kit #77552 보러 가기
cell-signaling-technology-cut-tag-assay-kit
  • CUT&Tag utilizes a secondary incubation step after the primary antibody incubation step to boost signal strength.
  • CUT&Tag is performed under high salt conditions. Both methods are compatible with histone analysis. However, CUT&Tag is less compatible with transcription factors and cofactors because high salt conditions can interfere with target protein-DNA interactions. This is particularly true for less abundant or weakly bound targets.
  • CUT&RUN uses Ca2+-activated pAG-MNase to cleave the DNA while CUT&Tag uses Mg2+-activated pAG-Tn5 to cleave the DNA. The Tn5 is charged with Illumina adaptors that are added to the chromatin during the cleavage process.
  • CUT&Tag permeabilizes both the cellular and nuclear membrane after tagmentation to completely solubilize the CUT&Tag DNA. This leads to the presence of both tagmented DNA and genomic DNA in the final DNA sample, making CUT&Tag DNA incompatible with qPCR. If qPCR is desired, we recommend performing CUT&RUN.
  • CUT&Tag skips the in vitro adaptor ligation step required for CUT&RUN, allowing you to skip directly to PCR amplification of your DNA library for sequencing–saving you precious time.
  • CUT&Tag’s time savings is cumulative, meaning you’ll see even more time savings as you scale up the number of samples you have.
 Cell Signaling Technology CUT&Tag

CUT&Tag delivers:

Faster time to results 1-2 days from cells to DNA library. CUT&Tag is 25% quicker than CUT&RUN due to streamlined library prep.
Low sample requirement ~40x less sample than ChIP and ChIP-Seq1
Low sequencing depth = sequencing cost savings Only ~2 million high-quality reads are required thanks to low background.
  1. Kaya-Okur HS, Wu SJ, Codomo CA, et al. CUT&Tag for efficient epigenomic profiling of small samples and single cells. Nat Commun. 2019;10(1):1930. Published 2019 Apr 29. doi:10.1038/s41467-019-09982-5

Both CUT&Tag and CUT&RUN help you unravel protein-DNA interactions when you are short on time and/or sample.

Use the table below to figure out which method is the right one for you!

  CUT&Tag CUT&RUN
Compatible with Histones ✔ ✔
Compatible with Transcription Factors Depends ✔
Compatible with Cofactors Depends ✔
Compatible with qPCR X ✔
Compatible with NG-seq ✔ ✔
DNA Library Prep In vivo In vitro
Cells to Library DNA 1-2 days 2-3 days
Low Cell ✔ ✔
Single Cell Amenable ✔ X
Sequencing Depth 2 M 3-5 M

CUT&Tag is compatible with next-generation sequencing (NGS) and is ideal for studying how histone modifications regulate chromatin binding when sample and time are limited. It can also be used to study transcription factors when the antibody used is validated specifically for CUT&Tag by you or a commercial vendor like CST.

Order the CST® CUT&Tag Assay Kit #77552 to get all the reagents you need for your CUT&Tag experiment or purchase just the reagents you need a la carte. All CUT&Tag reagents are stringently validated in-house to ensure you’ll get high-quality reagents every time. You can also choose from an ever-expanding list of CST CUT&Tag-validated antibodies.

CUT&Tag reagents give you the same high-quality data possible with CUT&RUN in half the library prep time.

Analyzing Histone Modifications with CUT&Tag

Generate equivalent data to ChIP-seq and CUT&RUN with the CUT&Tag Assay Kit #77552, CUT&Tag Dual Index Primers and PCR Master Mix for Illumina #47415, or a la carte products when studying histone modifications like Tri-Methyl-Histone H3 (Lys4) or Tri-Methyl-Histone H3 (Lys27). Go from cells to library DNA in 1-2 days with 100,000 starting cells.

 

Cell Signaling Technology CUT&Tag

Figure. ChIP-seq, CUT&RUN, or CUT&Tag assays were performed with HCT 116 cells and Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb #9751, using the SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005, CUT&RUN Assay Kit #86652, and CUT&Tag Assay Kit #77552, respectively. DNA libraries were prepared using the DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795 for ChIP-seq and CUT&RUN samples, and the CUT&Tag Dual Index Primers and PCR Master Mix for Illumina #47415 for CUT&Tag samples. The figure shows binding across chromosome 12 (upper), including GAPDH (lower), a known target gene of H3K4me3.

CST CUT&Tag 관련 제품 보러 가기
CST CUT&Tag Resource Center
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