Advanced Genomic Interaction Profiling Kit: CUT&Tag Assay with pAG-Tn5 for Illumina Sequencing
Innovative Insights into Genomic Interaction: pAG-Tn5 CUT&Tag Assay Kit for Illumina Sequencing
Elevate your research endeavors with the pAG-Tn5 CUT&Tag Assay Kit meticulously designed for DNA-protein interaction studies on the Illumina sequencing platform. This versatile library construction kit is not only tailored for mammalian cells but can also be applied to tissues or plant samples post-special treatment, expanding its utility across diverse biological contexts.
Key Advantages of pAG-Tn5 CUT&Tag:
- Overcomes limitations of traditional ChIP-Seq technology.
- Time-saving and user-friendly operation.
- Boasts a high signal-to-noise ratio.
- Requires a minimal starting amount of cells.
How it Works:
Harnessing the power of Tn5 transposase fused with Protein A/G, this kit facilitates binding to antibodies and precise DNA cutting in proximity to the target. Post-DNA recovery from cells, direct PCR amplification completes the library construction process, streamlining your workflow for seamless genomic exploration.
Quality Assurance:
Each component within the kit undergoes stringent quality control measures to ensure unparalleled product stability and reproducibility, providing you with the confidence to delve into your research with precision.
Unveil the Potential:
Designed explicitly for the Illumina sequencing platform, this kit serves as your gateway to cutting-edge genomics. Whether you are delving into Illumina sequencing or embarking on library construction, the pAG-Tn5 CUT&Tag Assay Kit is your trusted companion for pioneering discoveries.
Revolutionary Genomic Exploration: CUT&Tag Assay Kit for Illumina Sequencing
Embark on a pioneering journey into protein-DNA interactions with our CUT&Tag Assay Kit meticulously designed for Illumina's high-throughput sequencing platform. Leveraging the innovative Cleavage Under Target & Tagmentation (CleavageUnder Target & Tagmentation) technology, this kit marks a paradigm shift in the study of protein-DNA interactions.
Key Features:
- Utilizes Protein A/G fused transposase for precise targeting under antibody guidance, facilitating DNA cleavage near the target site.
- Introduces necessary splicing sequences for streamlined DNA extraction and PCR amplification, enabling direct library construction for sequencing.
Advantages Over Traditional CHE-SEQ:
- Low cell input requirements.
- Short experimental duration.
- High signal-to-noise ratio.
- Excellent repeatability.
Tailored Applications:
Especially crafted for research domains such as early embryonic development, stem cells, tumor studies, and epigenetics, this kit unlocks unparalleled insights into complex biological processes.
Quality Assurance:
Rigorous quality control measures ensure the stability and reproducibility of the library construction process, allowing you to explore genomics with utmost confidence.
Uncover the Future:
With a focus on advancing genomics, the CUT&Tag Assay Kit for Illumina Sequencing stands as a beacon for researchers seeking efficiency, precision, and breakthroughs in the intricate landscape of protein-DNA interactions.
Product Composition and Optimal Storage Environment
| Box | Component | 4T | 12T | Storage Conditions |
|---|---|---|---|---|
| I | pAG-Tn5 Transposome (4uM) | 4 µL | 12 µL | -20℃ |
| I | 5% Digitonin* | 64 µL | 192 µL | -20℃ |
| I | 1 M MgCl2 | 10 µL | 30 µL | -20℃ |
| I | 10×ConA Binding Buffer | 120 µL | 360 µL | -20℃ |
| I | 10×Wash Buffer | 1 mL | 1.5 mL*2 | -20℃ |
| I | 10×Dig-300 Buffer | 500 µL | 1.5 mL | -20℃ |
| I | Antibody Buffer（-） | 300 µL | 900 µL | -20℃ |
| I | Proteinase K (20 mg/mL) | 4 µL | 12 µL | -20℃ |
| I | 5×Stop Buffer | 100 µL | 300 µL | -20℃ |
| I | 2×PCR Mix for NGS | 150 µL | 450 µL | -20℃ |
| II | ConA Beads | 40 µL | 120 µL | 2-4℃ to avoid repeated freezing |
| II | DNA Clean Beads | 750 µL | 1.2 mL*2 | 2-4℃ to avoid repeated freezing |
Critical Considerations for Successful Experimental Design
Sample Preparation:
Binding Efficiency Test: Prior to the main experiment, assess the binding efficiency between cells and ConA magnetic beads. A high binding rate is crucial for enhancing the success of the experiment.
Handling Live Cells: When working with live cells, handle them appropriately based on cell type. For suspension cell lines, discard the upper culture medium after centrifugation. For adherent cells, collect cells post-trypsin digestion, centrifuge, and discard the upper medium.
Viability Testing: Stain cells with trypan blue for viability testing, aiming for viability over 90%. Gentle handling of cells during the experiment is vital to preserve viability, as poor cell growth or cell death can lead to altered protein-DNA binding states, potentially causing background noise.
Use of Nuclear Cells: Utilize nuclear cells for experiments and adapt nuclear extraction conditions based on tissue types.
Antibody Selection:
Establishing Control Groups: Establish both positive and negative control groups in the experiment. For positive controls, recommend histone proteins with high expression, while the negative control group should consist of normal secondary antibodies and transposons (without primary antibodies).
Preference for ChIP Level Antibodies: In the experiment, it is preferable to use antibodies at the ChIP level. If unavailable, antibodies suitable for immunofluorescence (IF) can be tested.
Secondary Antibody Selection: Select secondary antibodies with high Protein A/G affinity and without modifications to ensure optimal results in the experiment.