Global RNA Methylation Quantification by LC-MS/MS

While traditional RNA modification detection methods often face challenges in quantification accuracy, specificity, and cross-sample comparability, LC-MS/MS technology offers a robust solution with its antibody-free detection, absolute quantification capability, and exceptional precision. CD BioSciences provides a comprehensive global RNA methylation quantification service utilizing advanced LC-MS/MS platforms to deliver precise, reproducible, and absolute quantitative data on ribonucleotide modifications, empowering your epitranscriptomic research with reliable molecular insights.

Basic Principles of LC-MS/MS

The quantification of global RNA methylation by LC-MS/MS operates on the principle of direct, physical measurement of RNA building blocks. First, purified RNA is completely digested into its individual nucleosides by specific enzymes. This mixture is then injected into the liquid chromatography (LC) system, which meticulously separates each nucleoside (including modified ones like m6A, m5C, etc.) based on their chemical properties, effectively purifying them in time.

These separated analytes are then introduced into the tandem mass spectrometer (MS/MS). Here, they are first ionized and filtered by their mass-to-charge ratio (m/z) in the first mass analyzer. Selected ions are then fragmented in a collision cell, and specific, unique fragment ions for each modified nucleoside are monitored in the second mass analyzer. This multiple reaction monitoring (MRM) mode provides exceptional specificity and sensitivity. Finally, by comparing the signal intensity of each modified nucleoside to a calibration curve constructed from pure, known standards, the method achieves absolute quantification, providing precise molar ratios (e.g., fraction of m6A per total adenosine) in the original RNA sample.

Fig.1 LC-MS/MS-based characterization of the methylated positional isomers of adenosine originating from yeast mRNA. (Jora M, et al., 2019)

Features of LC-MS/MS

LC-MS/MS-based RNA methylation quantification service offers distinct advantages over other methodologies, providing researchers with precise, comprehensive, and reliable data to drive discovery in epitranscriptomics.

1. Gold Standard Method: Direct, label-free detection based on intrinsic physicochemical properties ensures exceptional specificity, free from antibody-related bias or variability.
2. True Absolute Quantification: Calibrated standard curves enable precise reporting of molar fractions or absolute amounts, allowing for reliable, quantitative cross-sample and cross-study comparisons.
3. High-Throughput Multiplexing: Simultaneously quantify a broad panel of RNA modifications in a single run for an efficient and holistic view of the epitranscriptomic landscape.
4. High Sensitivity & Accuracy: Exceptional sensitivity and reproducibility allow for the accurate measurement of low-abundance modifications and biologically significant subtle shifts, even from limited samples.

Our Services

As a leader in epigenetics research services, CD BioSciences offers comprehensive global RNA methylation quantification by LC-MS/MS to support your epitranscriptomic studies. Utilizing gold-standard liquid chromatography-tandem mass spectrometry (LC-MS/MS), we provide absolute quantification of key RNA modifications with high specificity, sensitivity, and reproducibility. Our end-to-end service covers expert sample preparation, rigorous quality control, multiplexed analysis, and detailed reporting with biological insights, enabling reliable cross-sample comparisons and accelerating discoveries in development, disease, and biomarker research.

Targeted RNA Modification

* The panel above represents our core detectable modifications. Please inquire about the quantification of additional specific RNA modifications.

Workflow of LC-MS/MS Service

Our Advantages

1. Flexible Sample Compatibility: Our optimized protocols support a wide range of sample types, including total RNA, mRNA, non-coding RNA, and RNA extracted from diverse sources such as cells, tissues, biofluids, and FFPE samples.
2. Commitment to Rigorous Quality: Stringent quality control is integrated at every stage, from sample acceptance to final data validation, guaranteeing high sensitivity, accuracy, and consistency even for low-abundance samples.
3. Customizable & Collaborative Approach: We work closely with you to tailor the service to your specific research needs, whether focusing on a core panel of modifications or developing methods for novel targets.
4. End-to-End Expertise: Your project is managed by our dedicated team of epigenetics specialists and analytical chemists, who oversee every step from experimental design to data interpretation, providing expert support throughout.

At CD BioSciences, we provide end-to-end LC-MS/MS-based RNA methylation quantification services that combine gold-standard methodology with deep scientific expertise. From flexible sample support and absolute multiplexed quantification to rigorous quality control and customized project design, we deliver the precise, reliable data you need to drive discovery forward. Contact our team today to discuss your specific research objectives and let us help you transform your epitranscriptomic insights into impactful results.

Reference

1. Jora M, Lobue P A, Ross R L, et al. Detection of ribonucleoside modifications by liquid chromatography coupled with mass spectrometry[J]. Biochimica et Biophysica Acta (BBA)-Gene Regulatory Mechanisms, 2019, 1862(3): 280-290.