| Description: | The EpiNext CUT&RUN RNA m6A-Seq Kit is a complete set of optimized buffers and reagents designed to enrich an RNA fragment containing m6A from low input RNA and to prepare a library for next generation sequencing using Illumina platforms such as Illumina Genome Analyzer II, HiSeq and MiSeq systems. The innovative working principle, optimized protocol, and components of the kit allow for the capture of the m6A fragment with minimal non-specific background levels. The enriched RNA is specifically suitable to construct both non-barcoded (singleplexed) and barcoded (multiplexed) libraries quickly, allowing m6A regions to be mapped with less bias and at a high resolution. This kit has the following advantages and features: High enrichment: Use RNA cleavage enzyme mix to simultaneously fragment RNA and cleave/remove any RNA sequences at both ends of the target m6A -containing sequences without affecting RNA regions occupied by the antibody. Short RNA fragments are generated only bound with anti- m6A antibody. True target m6A-enriched regions can, therefore, be reliably identified and high-resolution mapping achieved.Low Input: Unbound RNA cleavage and immunocapture are processed in the same single-tube, which enables the maximal protection of the target m6A-containing regions and the minimized sample loss, allowing the input RNA to be as low as 500 ng.Minimal Background: Cleavage of unbound RNA sequences in the two ends of the target m6A-containing sequences enables the minimized MeRIP/sequencing background, allowing data analysis with <10 million reads. Fast, streamlined procedure: The procedure from RNA to library cDNA is less than 6 hours with <1 h of hands-on time. Highly convenient: The kit contains all required components for each step of the CUT&RUN RNA m6A -Seq, which are sufficient for both m6A-containing RNA sequence capture and captured cDNA library preparation, thereby allowing CUT&RUN RNA m6A �Seq to be the most convenient with reliable and consistent results. |
| Further Information: | This kit contains all necessary reagents required for carrying out a successful m6A-Seq starting from total RNA. In the reaction, RNA sequences at both ends of the target m6A-containing regions are cleaved/removed and the target m6A-containing fragments are pulled down using a beads-bound m6A capture antibody. The target m6A-containing RNA fragments are then reverse-transcripted ligated with adaptors. The ligated RNA is released, purified, and amplified with a high-fidelity PCR mix for library cDNA construction. The cDNA is then cleaned, released, and eluted. Included in the kit is a negative control non-immune IgG, which can demonstrate the efficacy of the kit and performance at the enriched RNA quantification or bioanalyzer analysis step. |
