m6A-eCLIP kit

Highlights        

High Throughput and Robust Workflow
A simplified version of the Nature Methods-published eCLIP protocol enables profiling m6A marks with 40-fold less mRNA input than other approaches

Highly Reproducible with accurate data
Significant improvements in signal/noise reproducibly identifies m6A modified RNAs

Unbiased with high specificity
m6A eCLIP captures the DRACH motif and recapitulates other m6A profiling methods

The m6A eCLIP Kit is a targeted RNA modification kit that has applications in many fields of research. Starting with purified RNA, the kit allows researchers to globally map m6A RNA modification states, which has been associated with cancer, autoimmune disease, obesity, and other disease indications.

N6-methyladenosine (m6A) is the most prevalent epitranscriptomic RNA modification found in eukaryotes, and m6A has been shown to regulate many stages of RNA biology including splicing, secondary/tertiary structure, localization, stability, and translation. Adenosine modifications are actively added, recognized, and removed by a series of “writer”, “reader”, and “eraser” proteins, making m6A a dynamic and reversible system for regulating RNAs.

The m6A eCLIP Kit combines the use of an m6A antibody with the eCLIP technology, based on Van Nostrand et al. (Nature Methods, 2016) that allows user to accurately and robustly identify and map RBP binding sites on target RNAs. In pairing m6A and the eCLIP technology, RNA is chemically fragmented into 100 nucleotides or smaller fragments, followed by magnetic immunoprecipitation with a m6A antibody. After immunoprecipitation, isolated RNA fragments are subjected to library preparation including adapter ligation, reverse transcription, and PCR amplification. The streamlined 4-5 day protocol is easy to process and produces high quality libraries, and can be broken down into multiple start and stop steps. The kit enables the user to do mRNA isolation, m6A IP and library prep on up to 8 high quality samples.

Learn more about our eCLIP technology