Description
The kit is designed to prepare CUT&Tag libraries for mammalian cells for the Illumina high-throughput sequencing platform. The prepared libraries can be used for Single-end sequencing and Paired-end sequencing. Cleavage Under Target & Tagmentation (CUT&Tag) is a new method for studying protein interactions with DNA. Protein A/G-Tn5 transposase fusion protein and the target protein can form an immune complex through antibody bridging. Tn5 transposase is activated in the presence of Mg2+, and so short sequences containing adapters can be inserted into both ends of the DNA fragment that binds to the target protein. Finally, library can be constructed by PCR amplification. The kit is suitable for DNA research on the interaction of DNA-binding proteins such as transcription factors and co-regulators. Compared with traditional ChIP-seq, it has the advantages of more accurate and stable protein-DNA interaction information in low input of cells.
10-105 mammalian cells, or processed cell nucleus, and eukaryotic cells without cell wall (such as plant cell protoplasts, etc.).
Box 1 is stored at 2℃-8℃ for 1 year and Box 2 is stored at -18℃ or below for 1 year.
Dry ice (-70℃)
| Component | KP172-01 (12 rxns) | KP172-02 (48 rxns) | |
| Box1 | ConA Magnetic Beads II | 60 μl | 240 μl |
| CUT&Tag DNA Clean Beads | 2 ml | 4 ml × 2 | |
| Box2 | ConA Beads Binding Buffer | 660 μl × 2 | 2.7 ml × 2 |
| Cell Wash Buffer | 10 ml × 3 | 40 ml × 3 | |
| Nuclear Extraction Buffer | 6 ml | 6 ml × 4 | |
| 1×Pro-Wash Buffer | 10 ml | 40 ml | |
| pAG-Tn5 | 12 μl | 48 μl | |
| Tagmentation Buffer | 600 μl | 2.4 ml | |
| Protease Inhibitor | 420 μl | 840 μl × 2 | |
| Ab Enhancer | 20 μl | 80 μl | |
| Binding Enhancer | 80 μl | 320 μl | |
| 500 mM EDTA | 30 μl | 120 μl | |
| Lysis Enhancer | 20 μl | 80 μl | |
| Proteinase K（20 mg/ml） | 20 μl | 80 μl | |
| 2×CUT&Tag Library Amplification Mix | 300 μl | 600 μl × 2 | |
| Nuclease-free Water | 1 ml | 4 ml |
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