CD Genomics provides quantitative analysis of various tRNA modifications from total RNA samples based on LC-MS methods, to profile and characterize global modifications of tRNAs. We have the expertise to operate LC-MS/MS system and handle MS/MS data. This service contributes to a better understanding of tRNA biogenesis, structure, function, and implications in disease by delivering sensitivity, precision, accuracy, dynamic range of the quantification data.
Overview
Transfer RNA (tRNA) is identified as the most abundant RNA in a human cell and the most extensively modified RNA with an average of 13 modifications per molecule. In protein synthesis, tRNA serves as the adaptor of amino acids and the genetic code and carries a specific amino acid sequence to the ribosome for peptide extension. A tRNA is about 80 nucleotides long and contains an anticodon that base pairs with a complementary codon on mRNA. tRNA modification concentrates in two hotspots, the anticodon loop and tRNA core region, stabilizing the structure of tRNA.
Mass spectrometry (MS) is a powerful tool for comprehensive mapping of tRNA modifications and determination of the types of tRNA modifications and their extent. tRNA is first purified using affinity chromatography and digested with ribonucleases to generate oligonucleotides, followed by MS analysis. We provide LC-MS/MS service to profile and characterize tRNA modifications, enabling qualitative and quantitative analysis of tRNA modifications. Our service can determine the total tRNA modification content and condition-dependent tRNA modifications in response to stresses or diseases.
Features
Quantitative Analysis
High Coverage
Bioinformatics Analysis
Validated Process
Provide quantitative information on complex tRNA modifications
Simultaneous detection of 36 nucleoside modifications in tRNA
Data QC; profiling of tRNA modifications; quantification of
Validated experimental process using LC-MS/MS with MRM
Project Workflow
1. tRNA Isolation
Isolation of tRNA from total RNA sample using PAGE electrophoresis.
2. tRNA Digestion
Digest tRNAs down into single dephosphorylated nucleosides with ribonucleases.
3. LC-MS/MS
The nucleosides are subjected to LC-MS/MS analysis with MRM mode.
4. Data Analysis
Raw and normalized peak data; quantitative analysis; differential analysis.
Sample Requirements
We work with a wide range of sample types including protein solution, fresh tissue, cultured cells, blood, and microbial sample. Please feel free to contact us for sample size.
Sample Storage: extracted protein should be stored at -80°C. Avoid repeated freezing and thawing.
Shipping Method:When shipping the sample, it is stored in a 1.5 mL Eppendorf tube, sealed with a sealing film. Shipments are generally recommended to contain 5-10 pounds of dry ice per 24 hours.
Deliverable: data QC report, MS results, integrated experimental report (materials, methodologies, and bioinformatics analysis).
References:
Pan T. Modifications and functional genomics of human transfer RNA. Cell research, 2018, 28(4): 395-404.
Lorenz C, Lünse C E, Mörl M. tRNA modifications: impact on structure and thermal adaptation. Biomolecules, 2017, 7(2): 35.
* For Research Use Only. Not for use in diagnostic procedures.
