Description
The EpiNext™ CUT&RUN Fast Kit is a complete set of optimized reagents required for carrying out a successful CUT&RUN starting from mammalian cells or isolated nuclei/chromatin. The kit is designed to quickly enrich specific DNA complexes with a protein (histone or transcription factor) from low input cells/chromatin and to identify or map in vivo protein-DNA interaction by next generation sequencing using Illumina platforms or other methods such as qPCR. The innovative working principle, optimized protocol, and components of the kit allow for the capture of targeted protein/DNA complexes with minimized non-specific background levels. The captured DNA is specifically suitable for the construction of both non-barcoded (singleplexed) and barcoded (multiplexed) libraries to map target protein-DNA interaction regions with less bias and at a high resolution.
- High Enrichment: Uses a unique nucleic acid cleavage enzyme mix, which has low sequence bias, to simultaneously fragment chromatin and cleave/remove any DNA sequences in both ends of the target protein/DNA complex without affecting DNA occupied by the target protein. The enriched DNA >20 bps can be efficiently and quickly recovered. Thus, the target protein-enriched regions can be reliably achieved and identified at high-resolution mapping.
- Low Input Materials: Robust sonication-free fragmentation, unbound DNA cleavage, and immunocapture are all processed in the same single-tube with on-beads ligation. This method uses both cells and tissues and allows for maximal degradation protection of the target protein with minimal sample loss. As a result, the input cell amount can be as few as 500 cells, or chromatin amount can be as low as 0.1 µg.
- Minimal Background: Cleavage of unbound DNA sequences in the two (2) end of the target protein/DNA complex in situ enables the minimized immunocapture background, allowing the sequencing data analysis with <10 million reads.
- Fast, Streamlined Procedure: The procedure from cells to library DNA is less than 3 hours.
- Highly Convenient: The kit contains all required components for each step of the CUT&RUN, thereby allowing the EpiNext™ CUT&RUN Fast Kit to be the most convenient with reliable and consistent results.