Cleavage Under Targets and Tagmentation Service

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Cleavage Under Targets and Tagmentation Service

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CD BioSciences offers cleavage under targets and tagmentation technology service, based on extensive expertise in plant molecular biology, and provides clients with innovative solutions to accurately screen DNA that interacts with plant proteins with high-resolution and sensitivity equipment.

The Principle of Cleavage Under Targets and Tagmentation

Cleavage under targets and tagmentation is an emerging method for protein-DNA interaction studies. It works by causing the antibody-enzyme complex to attach to a protein target selectively, then cleaving the DNA region adjacent to the target. The extracted DNA is amplified to obtain a library and then sequenced. Recent studies have shown that cleavage under targets and tagmentation technology can be successfully applied in plants with better results than ChIP-seq.

Schematic diagram of the cleavage under targets and tagmentation experiment flow.  Figure 1. Schematic diagram of the cleavage under targets and tagmentation experiment flow. (Kaya-Okur, H. S., et al., 2020)

Service Content

Experimental content

  • Cell binding to magnetic beads, modulation of cell permeability.
  • Antibody and enzyme (VTn5 transposase) complex preparation.
  • Mg2+ activates transposon to fragment target DNA.
  • DNA fragment extraction, high-throughput sequencing.
  • Bioinformatics Analysis.
Experimental content.

Data analysis

Expansion services

  • Multi-cleavage under targets and tagmentation service

We also provide the multi-cleavage under targets and tagmentation service. Compared to traditional technology, it enables the simultaneous study of two targets in one sample without the use of secondary antibodies, reducing the process.

  • Optimization service of cleavage under targets and tagmentation product recovery

To improve recovery, we select the appropriate combination of antibodies and enzymes for clients to achieve optimal target specificity and lysis efficiency. Our optimization services include antibody concentration, incubation time, and temperature.

  • Optimization service of library construction methods

We offer optimized DNA purification methods to obtain high-quality DNA fragments. We utilize enzymatic reactions to generate junctions compatible with downstream sequencing platforms and carefully quality-check the libraries to ensure accurate and reliable sequencing results.

Notification for Sample Preparation and Handling

Sample type Details
Cells Highlights ≥ 4×105, transcription factor ≥ 106, cell viability ≥ 90% after resuscitation
Tissues ≥ 500mg
DNA library Total volum ≥ 25uL, concentration ≥ 2ng/uL, total amount ≥ 50ng, P7 and P5 index.
Antibodies The antibody (primary antibody) for the experiment needs to be provided by clients, it is used for Western Blot verification, the antibody should not be diluted, and should not be repeatedly frozen and thawed.

Deliverables

Cleavage under targets and tagmentation lab report

High-throughput sequencing raw data

Bioinformatics analysis report

Technology Advantages

  • Low sample starting volume, compatible with as few as 60 cells, even single cells.
  • ChIP-grade antibodies are not required.
  • Low background noise and high signal-to-noise ratio.
  • Good reproducibility between parallel samples.
  • Ultra-high activity, both pG/A & Tn5 activity, high DNA fragmentation activity.

CD BioSciences is a leading biological company in plant protein research. We are committed to providing reliable and comprehensive cleavage under targets and tagmentation analysis data to help clients unravel the complexities of plant protein function and chromatin dynamics. If you are interested in our service, please contact us for more details.

Reference

  1. Kaya-Okur, H. S., et al. (2020). Efficient low-cost chromatin profiling with cleavage under targets and tagmentation. Nature Protocols.15(10), 3264-3283.
  2. Tao, X., et al. (2020). Efficient chromatin profiling of H3K4me3 modification in cotton using cleavage under targets and tagmentation.Plant Methods. 16, 120.

For research use only, not for clinical use.