m1A RNA Enrichment & Quantification Kit

N¹-methyladenosine (m¹A), the methylation of adenosine in the N1 position, is a prevalent and reversible post-transcriptional RNA modification that can occur on messenger RNA (mRNA), transfer RNA (tRNA) and ribosomal RNA (rRNA). Recent studies have revealed the crucial role of m¹A 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 (MeRIP) is a popular method to observe and map the location of RNA modifications in specific genes as well as transcriptome-wide.

The RayBio^® m¹A (N¹-methyladenosine) RNA Enrichment and quantification kit (QMERIP-M1A) identifies the abundance and enrichment location of m¹A by MeRIP and quantitative real-time PCR. The enriched m¹A RNA could also be used for transcriptome-wide profiling of m¹A. In this assay, RNA sample is digested into fragments consisting of 100~500 nucleotides followed by incubation with an antibody against m¹A 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-M1A 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 m¹A 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. m¹A RNA immunoprecipitation (RIP).
4. qPCR assay for m¹A-RIP.
5. Calculate the relative m¹A 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.