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)
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.
Modification analysis of tRNAs from different sources.
tRNA is isolated from total RNA samples using electrophoresis.
tRNA is hydrolyzed into single dephosphorylated nucleosides with ribonuclease.
LC-MS/MS analysis of nucleosides.
Fig 2. Analysis of LC-MS data from modified ribonucleoside studies. (Dan, S.; et al. 2014)
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).
BOC Sciences supports HPLC-coupled mass spectrometry technology for the analysis and characterization of the global modification of tRNAs.
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