Sigma-Aldrich Magna Merip M6A Assay

N6-methyladenosine (m6A) represents an abundant RNA modification that is conserved across many different species, ranging from plants, to yeast, to mammals. Similar to the 5-methylcytosine (5-mc) modification of DNA, m6A is a reversible chemical modification of RNA. m6A RNA modifications are an emerging topic in controlling cell fate transitions in mammalian embryonic stem cells. m6A is one of the most prevalent modifications on both mRNAs and noncoding RNAs in eukaryotes with an estimated 12,000 m6A sites in over 7,000 genes. The mark is deposited by a heterodimer of methyltransferase-like 3 and 14 (Mettl3 and Mettl14) and can be removed by the RNA demethylase enzymes FTO and ALKBH5. Obesity risk gene FTO encodes the first identified m6A demethylase. Mutations in FTO have been associated with increased risk for obesity and type II diabetes. Recently, impairment of the status of m6A regulation has lead to prolonged expression of Nanog in ES cells and the inability to exit self-renewal stages toward differentiation.

Methylated (m6A) RNA immunoprecipitation (MeRIP) is a method to monitor the status of m6A and map the location of m6A RNA modifications transcriptome-wide. The Magna MeRIP M6A Kit uses the MeRIP method to enable identification and transcriptome-wide profiling of m6A RNA modifications. In the MeRIP assay, RNA is chemically fragmented into 100 nucleotides or smaller fragments followed by magnetic immunoprecipitation with a monoclonal antibody toward m6A. After immunoprecipitation, isolated RNA fragments can be subjected with qRT-PCR or RNA sequencing (MeRIP-seq). The streamlined protocol is easy to process and can produce results with high SN ratios.

The Magna MeRIP m6A Kit streamlines the MeRIP (Methylated RNA Immunoprecipitation) method to enable identification and transcriptome-wide profiling of m6A (N6-methyladenosine) RNA methylation sites. Suitable for MeRIP-Seq and qRT-PCR of methylated RNA transcripts.

RNA Fragmentation Buffer 10X (0.5 mL);0.5M EDTA (0.5 mL);IP Buffer 5X (15 mL);Magnetic Beads A/g Blend (315 μL);Anti-N6-methyladenosine (m6A), clone 17-3-4-1 (100 μg);RNase Inhibitor (90 μL);N6-Methyladenosine, 5′-monophosphate sodium salt (m6A) (10 mg);Normal Mouse IgG (125 μg);MeRIP Primers Human EEF1A1 Positive (75 μL);MeRIP Primers Human EEF1A1 Negative (75 μL)

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