CUT&Tag Sequencing

CUT&Tag (Cleavage Under Targets and Tagmentation) Sequencing is a next-generation sequencing-based method for the study of protein-DNA interactions. CUT&Tag is a new generation of ChIP-Seq without suffering from a number of practical and economical limitations, for it does not require cells to be lysed nor chromatin to be fractionated.

Overview

Compared with traditional ChIP-Seq methods, CUT&Tag eliminates the need for cross-linking, sonic interruption, end smoothing, and splicing. Therefore, CUT&Tag is time-efficient, requires less sample volume, has a low background signal, and is reproducible. Researchers can even use CUT&Tag for single-cell sequencing. Combined antibody-targeted cleavage induced by protein A-Tn5 fusion with high-throughput DNA sequencing, CUT&Tag identifies binding sites for DNA-associated proteins. Once the antibody binds to the target protein (transcription factor, histone, etc.), protein A binds directly to the antibody and carries a Tn5 transposase that specifically cuts the DNA fragment near the target protein and joins the DNA fragment, allowing for high-throughput sequencing after library construction. It can be used to precisely map the global DNA binding sites of any protein of interest. CUT&Tag is a prominent technology that can turn the study of protein-DNA interactions into a routine operation similar to PCR reactions, which will have revolutionary implications for the study of gene regulation, epigenetics, and many other fields.

Features

Low Sample Requirement & Efficient Genome-Wide Bioinformatics Analysis One-stop Service
Sensitive, efficient, repeatable, low background noise, and less sequencing depth is required. Precisely map the global DNA binding sites of any protein of interest. Our integrated bioinformatics pipeline can be tailored to suit your project. Provides one-stop service for library construction, sequencing, sample QC and data analysis.

Project Workflow

Sample Preparation

1. Sample Preparation

DNA purification;
quality assessment and quantification.

Library Preparation

2. Library Preparation

fragmentation;
library preparation.

Sequencing

3. Sequencing

Illumina HiSeq;
PE 50/75/100/150.

Data Analysis

4. Data Analysis

Visualize and preprocess results, and perform custom bioinformatics analysis.

CUT and Tag Sequencing

Bioinformatics Analysis Pipeline

In-depth data analysis:

  • Data quality control;
  • Spike-in alignment & scaling factor;
  • SEACR peak calling and Fragment proportion in Peaks regions;
  • Data visualization;
  • Differential analysis;
  • Additional alternatives.

Sample Requirements

Cell sample ≥ 50,000 cells; Tissue sample ≥ 5 mg

Sample Storage: Samples is recommended be frozen in liquid nitrogen and stored at -80°C.

Shipping Method: Samples are supposed to be stored in a 1.5 mL microcentrifuge tube, sealed with sealing film. Shipments are generally recommended to contain 5-10 pounds of dry ice per 24 hours.

Deliverable: FastQ, BAM, coverage summary, QC report, full statistical analysis & alignments, custom analysis reports on customer request.

References:

  1. Kaya-Okur HS, Janssens DH, Henikoff JG, et al. Efficient low-cost chromatin profiling with CUT&Tag. Nature Protocols. 2020:1-20.
  2. Kaya-Okur HS, Wu SJ, Codomo CA, et al. CUT&Tag for efficient epigenomic profiling of small samples and single cells. Nature communications. 2019;10(1):1-10.
* For Research Use Only. Not for use in diagnostic procedures.


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