m6A RNA Enrichment & Quantification Kit

N⁶-methyladensoine (m⁶A), the methylation of adenosine in the N6 position, is a prevalent and reversible post-transcriptional RNA modification that decorates mRNA, tRNA and rRNA. Recent studies have revealed the crucial roles of m⁶A RNA modification in multiple cellular processes, including RNA structural stability, folding, interactions with proteins, cell viability, impaired self-renewal ability, cell proliferation, and cell death. Methylated RNA immunoprecipitation (RIP) is a gold standard method to monitor the status and map the location of RNA modifications in specific genes and in transcriptome wide.

The RayBio^® m⁶A (N6-methyladenosine) RNA Enrichment and quantification kit (QMERIP-M6A) identifies the abundance and enrichment location of m⁶A by MeRIP and quantitative real-time PCR. The enriched m⁶A RNA could also be used for transcriptome-wide profiling of m⁶A. In this assay, RNA sample is digested into fragments consisting of 100~500 nucleotides followed by incubation with an antibody against m⁶A conjugated to a magnetic bead. Then, the enriched RNA is released, purified, and eluted for qRT-PCR analysis. Included in the kit is a non-immune IgG control conjugated to a magnetic bead and two pairs of primer controls for Hela cell RNA. QMERIP-M6A uses a fast and streamlined procedure so the assay can be finished in 5 hours with a hands-on time of less than 1 hour. This kit has been validated for human, mouse, and rat m⁶A enrichment.

• Fast, streamlined procedure, finished in 5 hours
• Hands-on time is less than 1 hour
• Enriched RNA for transcriptome-wide profiling

• Fluorescence PCR instrument and compatible PCR plate or tubes
• Sterile nuclease-free pipette tips (barrier tips recommended) and microfuge tubes
• Vortex mixer
• Magnetic separator
• Total RNA, mRNA, tRNA or rRNA
• Primers for target gene of interest, with or without probe

1. Prepare all reagents, samples and standards as instructed.
2. RNA digestion.
3. m6A RNA immunoprecipitation (DIP).
4. qPCR assay for m6A-RIP.
5. Calculate the relative m6A enrichment.

The entire kit can be stored at -20°C or -80°C for a period of 12 months prior to opening. After opening, the reagents are valid for at least 6 months. Avoid repeated freeze-thaw cycles.