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Arraystar - Go Beyond RNA
Small RNA Sequencing

tRNA Sequencing Service — Comprehensive tRNA Expression Profiling

Transfer RNAs are the most abundant small non-coding RNAs — and the hardest to profile accurately. Arraystar tRNA-seq uses the rtStar™ tRNA Pretreatment Kit to efficiently remove internal and terminal tRNA modifications, achieving exceptional tRNA-seq efficiency and accuracy for comprehensive tRNA expression profiling.

End-to-end sample-to-data service · First & only commercially available tRNA-seq

Overview

Accurate tRNA profiling starts with removing the modifications that block reverse transcription

Arraystar tRNA-seq is a sample-to-data service that accurately profiles tRNAs. Complete with sequencing data processing and a wealth of tRNA bioinformatic analysis, it is the first and only commercially available service of its kind — ready to spearhead your tRNA research in canonical and non-canonical tRNA biology.

The key to accurate tRNA sequencing is the rtStar™ tRNA Pretreatment Kit: tRNAs carry extensive internal and terminal post-transcriptional modifications that block reverse transcription and distort readout. By efficiently removing these modifications, the pretreatment delivers exceptional tRNA-seq efficiency and accuracy. Every project runs on performance-optimized tRNA sequencing methodologies with a rigorous QC process, supported by a comprehensive tRNA transcriptome reference for annotation.

What is tRNA-Seq?

tRNA sequencing (tRNA-seq) profiles transfer RNA expression at genome-wide scale. Because tRNAs carry dense post-transcriptional modifications that block reverse transcription, accurate tRNA-seq requires enzymatic pretreatment — such as Arraystar's rtStar™ Kit — before library construction.

Service at a Glance

Standard tRNA sequencing project — custom designs and add-on analyses available on request

Service NamePrice
tRNA Sequencing Service

Benefits

Why researchers choose Arraystar for tRNA sequencing

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rtStar™ Pretreatment

Efficiently removes internal and terminal modifications in tRNA to reach an exceptional level of tRNA-seq efficiency and accuracy.

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Optimized Methodologies

Performance-optimized tRNA sequencing methodologies with a rigorous QC process at every step.

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Comprehensive Reference

A comprehensive tRNA transcriptome reference supports full annotation of every detected tRNA.

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Thorough tRNA Annotation

Thorough tRNA annotation and bioinformatic analysis with publication quality graphics.

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Sample-to-Data Service

Sequencing data processing plus a wealth of tRNA bioinformatic analysis, with data visualization at publication quality graphics.

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First & Only of Its Kind

The first and only commercially available tRNA-seq service, spearheading canonical and non-canonical tRNA research.

Background

Why tRNA biology matters — and why it is hard to sequence

Transfer RNAs (tRNAs) are ubiquitous and the most abundant of all small non-coding RNA molecules. As a fundamental component in translation, tRNAs serve as the physical link between the mRNA coding and protein sequences. A wide variety of biological processes — such as cell proliferation [1], differentiation [1, 2], and apoptosis — are accompanied by changes in tRNA levels. Alterations of the tRNA repertoire can affect cell-fate choices during cell development (Fig. 1), and dysregulated tRNA repertoire can promote tumorigenesis and cancer progression.

tRNAs undergo by far the greatest number of, and the most chemically diverse, post-transcriptional modifications, which are essential for tRNA stability, folding, and decoding. As the amino acid carrier for peptide synthesis, tRNAs must also be charged with amino acids. However, these aminoacylated termini and the dense modification landscape block reverse transcription — which is why accurate tRNA-seq requires enzymatic pretreatment such as the rtStar™ tRNA Pretreatment Kit before library construction.

Figure 1. The effects of tRNA repertoire during cell fate determination
Figure 1. The effects of tRNA repertoire during cell fate determination. Changes in tRNA expression and modification reshape the translation landscape and contribute to cell proliferation, differentiation, and disease progression.

Background References

  1. Gingold H, et al. A dual program for translation regulation in cellular proliferation and differentiation. Cell, 2014. PMID: 25215487
  2. Pavon-Eternod M, et al. Overexpression of initiator methionine tRNA leads to global reprogramming of tRNA expression and increased proliferation. 2013. PMID: 23431330

tRNA Sequencing Workflow

Five phases from RNA sample to tRNA expression data

1

Sample QC

Total RNA quality and quantity assessment before the project proceeds.

2

rtStar™ Pretreatment

Enzymatic removal of internal and terminal tRNA modifications to unblock reverse transcription.

3

Library Construction

Adapter ligation, reverse transcription, and amplification with an optimized tRNA-seq protocol.

4

High-Throughput Sequencing

Sequencing with an optimized small RNA sequencing protocol for sensitive, quantitative detection of tRNA expression.

5

Bioinformatics

Expression profiling and differential expression analysis of the tRNA transcriptome.

Bioinformatics & Deliverables

A wealth of tRNA bioinformatic analyses — valuable for investigating tRNAs in biology and disease

Arraystar tRNA-seq includes a wealth of bioinformatics analyses, which are valuable for investigators to gain insights and better understand tRNAs in biology and disease. Every tRNA is annotated in detail with anticodon, sequence, dot-bracket structure notation, and expression levels (Fig. 1); differentially expressed tRNAs are presented with K-means clustering heatmaps, scatter plots, and volcano plots (Fig. 2); and expression levels of different tRNA types can be grouped by anticodon (Fig. 3, additional fee applies).

tRNA expression profiling annotated with anticodon, sequence, dot-bracket structure notation, and expression levels (panel 1)tRNA expression profiling annotation (panel 2)
Figure 1. tRNA Expression Profiling is annotated in detail with anticodon, sequence, dot-bracket structure notation, and expression levels.
Figure 2. Plots for differential expression analysis - K-means clustering heatmap, scatter plot, volcano plot
Figure 2. Plots for differential expression analysis. Differentially expressed tRNAs are presented on K-means clustering heatmap, scatter plot, and volcano plot.
Figure 3. Expression levels of different tRNA types grouped by anticodon
Figure 3. Expression levels of different tRNA types grouped by anticodon (additional fee applies).

Standard Deliverables

Research Applications

tRNA biology at the center of translation regulation — from development to disease

Translation Regulation & Cell Fate

tRNA repertoires shape translation programs during cell proliferation and differentiation — a dual program of translation regulation [1].

Cancer Biology

Threonine fuels glioblastoma through YRDC-mediated codon-biased translational reprogramming (Nature Cancer, 2024); SLFN12-mediated tRNA cleavage drives cancer cell death (Nature Chemical Biology, 2022).

Genetic & Neurological Disease

AAV-delivered suppressor tRNA overcomes a nonsense mutation in mice (Nature, 2022); neuronal Nsun2 deficiency produces tRNA epitranscriptomic alterations with synaptic and behavioral impacts (Nature Communications, 2021).

Protein Homeostasis & Mistranslation

Genetic interactions of tRNA-dependent mistranslation with FUS protein aggregates (Genes, 2023) and tRNAArg-derived fragments as arginine donors for protein arginylation (Cell Chemical Biology, 2020).

Mitochondrial & Codon-Biased Programs

Mitochondrial RNA polymerase PPR domain as an exoribonuclease for mtDNA replication (Nature Cell Biology, 2022); rare codon-based translational programs of cell proliferation (Genome Biology, 2020).

Anticodon Biology & Innate Immunity

Human SAMD9 as a poxvirus-activatable anticodon nuclease inhibiting codon-specific protein synthesis (Science Advances, 2023).

Sample Requirements

Official Arraystar sample submission requirements for tRNA sequencing projects

RNA Amount & Quality

  • Total RNA input: > 2 µg per sample (recommended minimum for the entire experiment including sample QC). Supply twice the minimum to avoid project delays.
  • Purification: TRIzol / RNA precipitation or an RNA isolation kit. Because tRNA is < 200 nt, use a kit specified to retain small RNAs (e.g. Qiagen miRNeasy).
  • Concentration: > 20 ng/µL by Nanodrop; OD260/280 ~2.0 (acceptable 1.7–2.1); OD260/230 > 1.8.
  • Integrity: sharp 18S/28S rRNA bands by gel, or RIN > 7.0 by Bioanalyzer (serum/plasma/exosome/FFPE RNA exempt).
  • DNase treatment: optional for gDNA removal; required if the sample is also used for qPCR.

Shipping Instructions

  • Ship RNA in nuclease-free water (> 20 ng/µL), freeze-dried, or in ethanol; store at −80 °C or in liquid nitrogen.
  • Use nuclease-free certified, screw-cap 1.5 mL microtubes; seal caps with Parafilm; place tubes in a plastic bag.
  • Use 10 kg dry ice as refrigerant; include a signed Project Form in a waterproof bag, separate from samples.
  • Avoid Thursday/Friday drop-offs before weekends or holidays; email the tracking number to your Arraystar representative.
Sample TypeRequirementNotes
Total RNA> 2 µg*tRNA-seq recommended minimum; small-RNA-retaining purification kit required
Cultured cells2 × 10⁶ cellsLyse immediately in TRIzol (1 mL per 5–10 × 10⁶ suspension cells or 10 cm² dish); ship on dry ice
Tissue10–25 mgRNAlater (≥10 volumes, 2–8 °C overnight), TRIzol (1 mL per 10–25 mg), or fresh-frozen in liquid nitrogen
Whole blood2–3 mL (0.3 mL/aliquot)tRNA-seq requires pretreatment → use the “With IP or Pretreatment” volumes; ship on dry ice
Plasma / Serum2–5 mL (0.3 mL/aliquot)EDTA or citrate anticoagulant; do NOT use heparin; ship on dry ice
ExosomesCustomer-isolatedResuspend in 200 µL PBS + 800 µL TRIzol (v:v = 1:4); ship on dry ice

* Recommended minimum per sample for the entire experiment in a single attempt, including sample QC; supply twice the recommended minimum to avoid project delays. If the minimum amount is not obtainable, contact support@arraystar.com for special arrangements. Arraystar performs sample QC upon receipt; QC assessment is final. Low amount/quality samples may proceed with consent — data quality and success rate may decline. Shipping address: ATTN: Samples Receiving (Project#______), Arraystar Inc., 9430 Key West Avenue #128, Rockville, MD 20850 USA, Tel: 888-416-6343.

FAQ

Common questions about tRNA sequencing

What sample types are compatible with tRNA sequencing?
We accept total RNA extracted from cultured cells, tissues, whole blood, plasma, serum, or other biological sources. Purified RNA should be intact with an OD260/280 of ~2.0 (acceptable 1.7–2.1), OD260/230 above 1.8, and a RIN above 7.0 where applicable. If you're unsure whether your sample type is suitable, please contact us for a free evaluation.
What is the minimum amount of RNA required?
We recommend more than 2 µg of total RNA per sample — the official Arraystar recommended minimum for the entire experiment in a single attempt, including sample QC. Supplying twice the recommended minimum helps avoid project delays. RNA integrity must be preserved, because degraded RNA cannot be rescued by downstream steps.
Why is tRNA pretreatment necessary before sequencing?
tRNAs carry extensive internal and terminal post-transcriptional modifications that block reverse transcription and distort readout, causing biased and inaccurate profiling. Arraystar's rtStar™ tRNA Pretreatment Kit enzymatically removes these modifications before library construction, enabling exceptional tRNA-seq efficiency, accuracy, and reproducibility across replicates.
Can tRNA-seq detect tRNA modifications?
No — this service profiles tRNA expression rather than modification status. For modification analysis, Arraystar offers dedicated services: tRNA Modification Sequencing (m1A, m3C, m1G, m2,2G), m7G TRAC-Seq and m3C HAC-Seq at single-nucleotide resolution, and LC-MS based tRNA modification analysis. These dedicated services complement tRNA-seq within Arraystar's broad tRNA research portfolio.
What kind of data will I receive?
You'll receive raw sequencing data (FASTQ), sample and sequencing QC reports, and tRNA expression tables annotated with anticodon, sequence, dot-bracket structure notation, and expression levels. Results include differential expression analysis with K-means clustering heatmap, scatter, and volcano plots, and expression levels of different tRNA types grouped by anticodon (additional fee applies).
How long does the service take?
Turnaround time typically depends on sample type, the number of samples, and project complexity, including any add-on bioinformatics analyses. We provide a firm timeline when your project is quoted. Please contact us directly for a time estimate based on your specific needs.

Selected Publications

Featured Client Publications in tRNA Research

  1. Wu X, et al. Threonine fuels glioblastoma through YRDC-mediated codon-biased translational reprogramming. Nature Cancer, 2024. PMID: 38519786
  2. Zhang F, et al. Human SAMD9 is a poxvirus-activatable anticodon nuclease inhibiting codon-specific protein synthesis. Science Advances, 2023. PMID: 37285440
  3. Lant JT, et al. Genetic interaction of tRNA-dependent mistranslation with fused in sarcoma protein aggregates. Genes, 2023. PMID: 36833445
  4. Wang J, et al. AAV-delivered suppressor tRNA overcomes a nonsense mutation in mice. Nature, 2022. PMID: 35322228
  5. Lee S, et al. Velcrin-induced selective cleavage of tRNALeu(TAA) by SLFN12 causes cancer cell death. Nature Chemical Biology, 2022. PMID: 36302897
  6. Liu Y, et al. The PPR domain of mitochondrial RNA polymerase is an exoribonuclease required for mtDNA replication in Drosophila melanogaster. Nature Cell Biology, 2022. PMID: 35449456
  7. Blaze J, et al. Neuronal Nsun2 deficiency produces tRNA epitranscriptomic alterations and proteomic shifts impacting synaptic signaling and behavior. Nature Communications, 2021. PMID: 34880236
  8. Guimaraes JC, et al. A rare codon-based translational program of cell proliferation. Genome Biology, 2020. PMID: 32102681
  9. Avcilar-Kucukgoze I, et al. tRNAArg-derived fragments can serve as arginine donors for protein arginylation. Cell Chemical Biology, 2020. PMID: 32553119

Ready to Profile the tRNA Transcriptome Accurately?

Arraystar tRNA-seq combines rtStar™ pretreatment with optimized tRNA sequencing methodologies and comprehensive bioinformatics — get a quote and a project timeline tailored to your study.