N6-methyladenosine (m6A) can be measured four main ways — competitive ELISA, dot blot, MeRIP-qPCR and LC-MS/MS — and the right one depends on whether you need a global level, a relative comparison, gene-specific enrichment or absolute quantification. m6A is the most abundant internal modification of eukaryotic mRNA and regulates RNA stability, splicing and translation, so it now appears in cancer, metabolism, neuroscience and virology projects. This guide compares the methods side by side and shows which RayBiotech kit fits each question.
Looking for a kit? Compare RayBiotech m6A ELISA, dot blot and MeRIP-qPCR kits.
Which m6A method answers which question?
| Method | What it tells you | Output | Typical sample | Strengths | Limits |
|---|---|---|---|---|---|
| Competitive ELISA (EIA) | Amount of m6A in a sample | Quantitative, from a standard curve | Per kit specification (RayBiotech EIA-m6A: plasma, serum, urine) | 96-well throughput, plate reader only | No gene- or site-level information |
| Dot blot | Relative global m6A between samples | Semi-quantitative | Extracted RNA; serum/plasma per datasheet | Fast, cheap, good first screen | Relative only; depends on equal loading |
| MeRIP-qPCR | m6A enrichment on specific transcripts or regions | Quantitative (% input / fold enrichment) | Extracted RNA | Gene-specific; follows up sequencing hits | Antibody-dependent; region-level, not single-base |
| MeRIP-seq / m6A-seq | Transcriptome-wide m6A map | Peaks across the transcriptome | Extracted RNA (relatively large input) | Discovery of modified transcripts | Cost, bioinformatics, ~100–200 nt resolution |
| LC-MS/MS | Absolute m6A/A ratio in digested RNA | Absolute quantification | Purified, nuclease-digested RNA | Gold-standard accuracy | Needs mass spectrometry and expertise |
A practical decision path
- “Does my treatment or knockdown change m6A overall?” Start with a dot blot on equal amounts of RNA, or an ELISA if your samples and the kit’s validated sample types match. Confirm a striking change by LC-MS/MS if you have access.
- “Is m6A on my gene of interest changed?” Use MeRIP-qPCR with primers on the region you expect to be modified (often near the stop codon or 3′ UTR), plus a known positive and negative control transcript.
- “Which transcripts are modified?” Use MeRIP-seq for discovery, then validate selected targets by MeRIP-qPCR.
- “Exactly where, at single-base level?” Use single-nucleotide methods such as miCLIP.
Tips that save experiments
- Start with clean RNA. Genomic DNA and degraded RNA distort dot blots and MeRIP. Check purity and integrity first — see A260/A280 for RNA and what degraded RNA looks like on a gel.
- Load equal amounts and stain the membrane (for example with methylene blue) for dot blots, so differences reflect m6A rather than loading.
- Include controls for MeRIP: an IgG pull-down, input RNA, and transcripts known to be highly and poorly modified.
- Match the kit to the sample. ELISA kits are validated for specific sample types; use them within the datasheet’s scope.
RayBiotech m6A kits
| Catalog # | Kit | Method | Output | Species | Compatible samples | Size |
|---|---|---|---|---|---|---|
| EIA-m6A | m6A (N6-methyladenosine) ELISA Kit | Competitive ELISA, colorimetric, 96-well microplate | Quantitative; sensitivity 0.435 ng/ml, detection range 0.435–10,000 ng/ml | Human, Mouse, Rat | Plasma, Serum, Urine | 1, 2 or 5 plate kit |
| DB-m6A | m6A (N6-methyladenosine) Dot Blot Assay Kit | Dot blot on nitrocellulose, chemiluminescent detection | Semi-quantitative; ≥48 samples per kit | Human, Mouse, Rat | Plasma, Serum, Extracted RNA | 1 kit |
| QMERIP-M6A | m6A RNA Enrichment & Quantification Kit | MeRIP (anti-m6A magnetic beads) + qRT-PCR; RNA fragmented to 100–500 nt; IgG and primer controls included | Quantitative; about 5 h, <1 h hands-on | Human, Mouse, Rat | Extracted RNA | 48 or 96 tests |
| DB-4-RNA | 4-RNA Methylation Dot Blot Combination Kit (m6A, m1A, m5C, m7G) | Dot blot | Semi-quantitative | Human, Mouse, Rat | Plasma, Serum, Extracted RNA | 1 kit |
Specifications as listed by RayBiotech for each catalog number; follow the manual shipped with the kit.
Request a quote for any of these kits, or send us your sample type and question and we will suggest the method that fits. Research use only.
Common questions
What is the best way to quantify m6A?
For absolute quantification, LC-MS/MS of digested RNA is the reference method. For routine comparisons across many samples, ELISA or dot blot is faster; for gene-specific changes, MeRIP-qPCR.
Is a dot blot quantitative?
No, it is semi-quantitative. It compares relative global m6A between samples loaded in equal amounts on the same membrane.
Can I measure m6A on a single gene with ELISA?
No. ELISA and dot blot report bulk m6A. To look at a specific transcript, enrich modified RNA by MeRIP and quantify your target by qPCR.
Which samples work with the RayBiotech m6A ELISA?
According to the datasheet, EIA-m6A is validated for plasma, serum and urine from human, mouse and rat. Check the kit manual before using other sample types.
Key method papers
Dominissini D, Moshitch-Moshkovitz S, Schwartz S, et al. Topology of the human and mouse m6A RNA methylomes revealed by m6A-seq. Nature. 2012;485:201–206. doi:10.1038/nature11112
Meyer KD, Saletore Y, Zumbo P, et al. Comprehensive analysis of mRNA methylation reveals enrichment in 3′ UTRs and near stop codons. Cell. 2012;149:1635–1646. doi:10.1016/j.cell.2012.05.003
Jia G, Fu Y, Zhao X, et al. N6-methyladenosine in nuclear RNA is a major substrate of the obesity-associated FTO. Nature Chemical Biology. 2011;7:885–887. doi:10.1038/nchembio.687
Linder B, Grozhik AV, Olarerin-George AO, et al. Single-nucleotide-resolution mapping of m6A and m6Am throughout the transcriptome. Nature Methods. 2015;12:767–772. doi:10.1038/nmeth.3453