Epitranscriptomic RNA modifications (e.g., m6A, 5-mC) are key genetic regulators linked to diseases like cancer. Creative Biolabs' RNA Methylation Assay Service offers m6A/5-mC quantification and single-nucleotide resolution mapping. It overcomes epitranscriptomic research bottlenecks, delivers high-resolution data from low-input samples, and provides validated, statistically confident insights to accelerate biomarker discovery and therapeutic development.
m6A and 5-mC modifications are dynamically regulated by specialized "writer," "eraser," and "reader" proteins. This dynamic state allows the marks to act as real-time sensors of cellular conditions. Measuring changes in m6A and 5-mC abundance or location (Differentially Methylated Regions, DMRs) provides profound insight into disease etiology, making these modifications compelling targets for epigenetic drug development and personalized medicine.
Fig.1 The chemical structure of methylated nucleotides. 5-methylcytosine (m5C), N6-methyladenosine (m6A), 2'-O-dimethyladenosine (m6Am), N1-methyladenosine (m1A), and N7-methylguanosine (m7G).1,3
The choice of methodology dictates the utility and resolution of the resulting data. We utilize a tiered approach:
Primarily for high-throughput screening or initial assessment, typically via ELISA-based colorimetric/fluorometric assays. Fast, suitable for various species, and provides total m6A or 5-mC abundance.
For the highest resolution and discovery, employing next-generation sequencing techniques:
Fig.2 The methods for methylation labeling localization include Elisa, MeRIP-seq, miCLIP, SELECT, and MazF methods.1,3
Our workflow ensures that every stage, from initial consultation to final report, maximizes data quality, statistical power, and budget efficiency, making the process clear and reliable for potential clients.
| Key Steps Involved | Activities and Expected Outcomes |
|---|---|
| Strategic Design & Power Analysis | Our experts collaborate with your team to review pilot data, define effect size, and apply simulation-based statistical frameworks (akin to the powerful magpie model) |
| Sample Processing & Quality Control | Client-provided RNA is rigorously assessed for quantity (down to 10 ng) and integrity (RIN). We utilize proprietary, low-input protocols, including optimized MeRIP/miCLIP variants, to preserve sample material. |
| High-Resolution Sequencing | Library preparation followed by next-generation sequencing (NGS). We prioritize techniques with single-nucleotide resolution and long-read compatibility to minimize ambiguity and capture isoform-specific methylation. |
| Advanced Bioinformatics & DMR Calling | Proprietary analysis pipelines are applied, including alignment, peak calling, and Differential Methylated Region (DMR) detection. We apply comprehensive metrics, including False Discovery Cost (FDC), for data filtering. |
| Comprehensive Biological Interpretation | Integration of methylation data with existing transcriptomic profiles (if available) for pathway analysis, functional annotation, and mechanism hypothesis generation. |
Required Starting Materials:
Final Deliverables:
Estimated Timeframe:
The typical timeframe for this service ranges from 8 to 12 weeks, depending on the complexity of the starting material, the required sequencing depth, and the scale of the custom bioinformatics required for the project scope.
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Creative Biolabs is dedicated to providing superior, customized solutions that elevate your epitranscriptomic research above standard service offerings. We combine strategic planning with technical flexibility to guarantee scientific excellence.
Customized End-to-End Epitranscriptome Solutions
Offer a comprehensive one-stop service including strategic study design, custom m6A and 5-mC library preparation, high-throughput sequencing, and functional biological interpretation, tailored to your discovery phase.
Integrated Statistical Quality-by-Design (QbD)
Upfront Strategic Design & Power Analysis guarantees project robustness, selecting optimal sample size and sequencing depth to meet statistical confidence targets (e.g., 90% power) and maximize budget efficiency.
Maximum Sample Flexibility and Low-Input Assurance
Deploy proprietary protocols optimized for precious clinical and scarce research samples, reliably processing Total RNA as low as 10 ng for global quantification and high-resolution mapping.
Unrivaled Custom Resolution and Technology Options
Provide flexible detection technology choices, including optimized MeRIP-seq/miCLIP variants for transcriptome-wide DMR mapping and Nanopore/SMRT compatibility for single-nucleotide resolution requirements.
Tiered Assay Options for Every Project Phase
Select the most appropriate assay based on resolution, budget, and project phase—from high-throughput ELISA-based global quantification for screening to full transcriptome-wide sequencing and functional annotation for validation.
Actionable Bioinformatics Deliverables
Advanced custom analysis pipelines deliver statistically filtered DMR lists, comprehensive QC metrics, and detailed Biological Pathway Analysis for immediate therapeutic or biomarker application, accelerating data-to-patentable insights.
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This case proposes a comprehensive power assessment method called magpie (m6A Granular Differential Analysis Power Inference): it first learns the characteristics of real data and synthesizes simulated data. During simulation, it can adjust sample size, sequencing depth, and effect size, and evaluate epitranscriptomic study designs using multiple metrics, including sensitivity, specificity, precision, and false discovery rate. Its core workflow consists of two independent simulation steps: preprocessing MeRIP-seq data to obtain read counts in candidate regions, then simulating data with the Gamma-Poisson model, and finally completing the assessment of DMR detection power and error rates.
Fig.3 A brief step for Magpie to function.2,3
Standard RNA-seq only measures gene expression (abundance), not the post-transcriptional chemical modification itself. You need a dedicated RNA Methylation Assay Service to specifically isolate and map the modified RNA fragments (m6A or 5-mC). Our service ensures you capture this critical regulatory layer that gene expression alone will miss.
We specialize in low-input research. While traditional methods require micrograms of RNA, our optimized protocols are validated to deliver high-quality mapping from as little as 10 ng of total RNA. This sensitivity makes studies on biopsies or circulating tumor cells feasible.
The issue is often inadequate statistical power. We address this upfront with our Integrated Study Design and Power Analysis. Before sequencing, we use advanced simulations to calculate the precise sample size and sequencing depth needed to achieve statistical significance, preventing the waste of time and resources associated with false negatives.
ELISA kits only provide a global percentage of m6A in the entire RNA pool. Our mapping service identifies the exact location (single-nucleotide resolution) of the methylation on specific transcripts. If you need to find a therapeutic target or diagnostic biomarker, you must know which transcript is modified, which requires our high-resolution mapping service.
Our service includes a full bioinformatics pipeline, taking you from raw data to a publication-ready report. We deliver not just files, but a detailed Biological Pathway Analysis and interpretation. We also provide ongoing consultation to help you understand the identified DMRs and their functional implications for your project.
Creative Biolabs is your strategic partner for unlocking the epitranscriptome. We provide a fully integrated solution that combines low-input, high-resolution sequencing with rigorous biostatistical design for m6A and 5-mC analysis. Our expertise ensures your project is scientifically sound, statistically confident, and poised for groundbreaking discovery.
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