tRNA Modification Analysis Services

RNA modification has gradually become a research hotspot. RNA contains various epigenetic modifications, and these epigenetic modifications can regulate RNA stability and translation to control gene expression. In addition, these modification plays an important role in tissue development and tumorigenesis. Post-transcriptional modifications are essential determinants of RNA quality control, and these modifications profoundly affect their structures and functions. Compared to mRNAs, tRNAs are more abundantly modified. Studies have showed that tRNA modifications have been linked to cancer and a series of diseases. For example, abnormalities in multiple modifications that occur on tRNAs may lead to birth defects and neurodevelopmental abnormalities in patients. Therefore, systematic study of tRNA modifications is key to understanding the relationship between disease and various modifications. In recent years, scientists have developed various tools and strategies to identify and quantify different modifications. Mass spectrometry (MS) is a powerful tool that can be used to comprehensively map tRNA modifications. In the analysis process, tRNAs are purified and digested to generate oligonucleotides, which are then analyzed by MS.

Fig 1. Workflow for the quantification of tRNA modification. (Dan, S.; et al. 2014)

Applications of tRNA Modification Analysis

• Extensive exploration and quantification of ribonucleosides.
• Facilitate the homeostatic studies of tRNA modifications in complex biological samples.
• Support the subsequent analysis of individual tRNA species and tRNA copy numbers.
• Disease research for the promising treatment.

Our Services

• LC-MS

BOC Sciences' LC-MS analysis service provides a complete solution from sample preparation to data analysis. Our experts simultaneously analyze more than 50 nucleoside modifications.

Workflow of Our LC-MS Analysis

• Preparation of tissue, cultured cells, blood, and microbial sample

Modification analysis of tRNAs from different sources.

• tRNA isolation

tRNA is isolated from total RNA samples using electrophoresis.

• tRNA digestion

tRNA is hydrolyzed into single dephosphorylated nucleosides with ribonuclease.

• LC-MS/MS

LC-MS/MS analysis of nucleosides.

• Bioinformatics data analysis

Fig 2. Analysis of LC-MS data from modified ribonucleoside studies. (Dan, S.; et al. 2014)

• UPLC-MS

We provide UPLC-MS analysis services to meet the needs of detailed analysis of complex biological samples. An octadecyl carbon chain (C18)-bonded silica matrix is employed for the rapid and efficient separation of 50 modified ribonucleosides, including positional isomers. The flexibility of our approach supports the detection and relative quantification of tRNAs using stable isotope labeling and multiple reaction monitoring (MRM).

Advantages of Our UPLC-MS Services

• Reduced time for the separation of modified ribonucleosides.
• Reliable identification of tRNA modification together with the unique fingerprint ionization pattern.
• Suitable for detection of multiple tRNA modifications, especially positional isomers.

• HPLC-MS

BOC Sciences supports HPLC-coupled mass spectrometry technology for the analysis and characterization of the global modification of tRNAs.

The Protocol of HPLC-MS Analysis

• RNA purification and quantification.
• Hydrolysis of tRNA into individual ribonucleosides.
• Reverse-phase HPLC separation.
• Identification and quantification of ribonucleosides by tandem quadrupole mass spectrometry.
• Multivariate statistical analysis of patterns of changes in the modified ribonucleoside set induced by the stress or stimulation.

Features of Our tRNA Modification Services

• Suitable for analyzing a variety of tRNA modification patterns.
• Applied to studies in bacteria, yeast and human cells.
• Accurate quantification of complex tRNA modifications from different types of RNA samples.
• Optimized experimental procedures and professional instrument operation knowledge.
• Highly sensitive and quantitative over a broad detection range, capable of simultaneously analyzing more than 50 nucleoside modifications in tRNA.
• State-of-the-art systems enable to deliver robust quantification results.