This product is developed for illumina high-throughput sequencing platform based on the principle of CUT & Tag (Cleavage Under Targets and Tagmentation) technology to study protein-DNA interactions. Compared with the traditional ChIP-seq technology, this product does not need to interrupt the traditional ultrasound, connectors and other cumbersome steps, the operation is more simple, with a short experimental cycle, low cell input, low antibody input, high library yield, high signal-to-noise ratio, suitable for embryonic development, stem cells, tumorigenesis, and other epigenetic fields of research.
CUT & Tag is a novel method to study DNA-protein interactions. It uses Tn5 transposase fused with Protein A/G, which precisely binds to the antibody under the guidance of Protein A/G, enabling the Tn5 transposon to target to the target site for enzyme cleavage, and at the same time, adding part of the junctions necessary for second-generation sequencing to the cleaved DNA, which can be extracted, amplified by PCR and purified to obtain a library for sequencing. After DNA extraction, PCR amplification and purification, the library can be obtained for sequencing, and the DNA information of the target site can be obtained after sequencing.
The reaction system of this product has been carefully optimized, all reagents in the experiment should be used as provided in this product, it is not recommended to change any reaction components or replace the components in this product with other equivalent products in order to avoid obtaining bad experimental results. If replacement is needed, please verify it first.
1. Suitable for illumina sequencing platform: The illumina connector is integrated and the constructed DNA library can be directly sequenced by NGS.
2. Wide cell range compatibility: from 100~105 A starting volume of cells was experimented with.
3. High signal-to-noise ratio: low background noise signal, good experimental repeatability.
4. Simple operation: the whole process of magnetic bead extraction and purification, simple and rapid operation.
5. pA/G-Transposome Mix has strong performance: pA/G structural domain has broad affinity for antibodies from rabbit, mouse and other sources, with precise targeting; optimized Tn5 enzyme can efficiently and accurately cut target DNA.
6. High quality sequencing: optimized PCR amplification system, high quality sequencing data.
Package 3-1 is composed as follows (stored at -80°C):
| individual parts making up a compound | AG12563 | AG12555 | AG12556 |
| pA/G-Transposome Mix | 4 μl | 12 μl | 48 μl |
Package 3-2 is composed as follows (stored at 4°C):
| individual parts making up a compound | AG12563 | AG12555 | AG12556 |
| ConA Beads Pro | 20 μl | 60 μl | 240 μl |
| DNA Clean Beads | 660 μl | 1 ml x 2 pcs | 8 ml |
| 10% SDS | 8 μl | 24 μl | 96 μl |
Package 3-3 is organized as follows (stored at -20°C):
| individual parts making up a compound | AG12563 | AG12555 | AG12556 |
| Lysis Buffer | 400 μl | 1.2 ml | 1.2 ml X 4 pcs |
| 10X ConA Binding Buffer | 100 μl | 300 μl | 1.2 ml |
| 10X Wash Buffer | 1 ml | 1.6 ml x 2 pcs | 1.6 ml X 6 pcs |
| 5% Digitonin Solution | 28 μl | 80 μl | 320 μl |
| 10X Dig-300 Buffer | 220 μl | 660 μl | 1.2 ml X 2 pcs |
| BE Buffer | 14 μl | 42 μl | 168 μl |
| MgCl2 Solution | 8 μl | 24 μl | 96 μl |
| Stop Buffer | 6 μl | 18 μl | 72 μl |
| AccuNext PCR Mix Ⅱ | 80 μl | 240 μl | 960 μl |
| Proteinase K (20 mg/ml) | 4 μl | 12 μl | 48 μl |
Attention:AccuNext Transposome building adaptor primers (Illumina) (Code No. AG12532 / AG12533 / AG12534) are required for the experiments, but are not prepared in this product and need to be purchased separately.AccuNext Information on the transposome building adaptor primers (Illumina) is provided in Appendix 1.
Package 3-2 4°C Storage (avoid freezing)
Package 3-3 -20°C Storage
Transportation Temperature: Package 3-1 Dry Ice Transportation
Package 3-2 Ice Pack Transportation
Package 3-3 Dry Ice or -20°C Ice Pack Transportation
