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tRNA Research Tools

rtStar™ tRNA Pretreatment & First-Strand cDNA Synthesis Kit — Remove tRNA Modification Barriers for Accurate cDNA Synthesis

The rtStar™ tRNA Pretreatment & First-Strand cDNA Synthesis Kit is designed for reverse transcribing tRNAs efficiently. It removes internal modifications of tRNAs to ensure the proceeding of reverse transcriptase during cDNA synthesis, delivering magnitudes of increase in tRNA detection sensitivity and quantification accuracy.

Efficient tRNA demethylation · for qPCR arrays, primer sets, and sequencing

Overview

The key to accurate tRNA reverse transcription

tRNAs carry extensive internal post-transcriptional modifications that block reverse transcriptase during cDNA synthesis. The rtStar™ tRNA Pretreatment & First-Strand cDNA Synthesis Kit efficiently removes internal modifications of tRNAs, whereas regular cDNA synthesis methods cannot, ensuring efficient first-strand cDNA synthesis.

This kit is the recommended companion for the nrStar™ tRNA PCR Array (Cat# AS-NR-001) and rtStar™ Pre-designed tRNA Primer Sets, and delivers magnitudes of increase in tRNA detection sensitivity, quantification accuracy, and assay discriminating power with the treated compared with untreated samples.

What is tRNA pretreatment?

tRNA pretreatment enzymatically removes the extensive internal post-transcriptional modifications that block reverse transcription during cDNA synthesis. The rtStar™ kit combines this pretreatment with first-strand cDNA synthesis, enabling efficient, accurate, and sensitive tRNA detection by qPCR and sequencing.

Service at a Glance

Standard catalog product

Product NameCatalog No.SizePrice
rtStar™ tRNA Pretreatment & First-Strand cDNA Synthesis KitAS-FS-00412 reactions

Benefits

Key advantages of the rtStar™ kit

⚗️

Efficient Modification Removal

Efficiently removes internal modifications of tRNAs to ensure the proceeding of reverse transcriptase during cDNA synthesis, whereas regular cDNA synthesis methods do not.

📈

Dramatically Improved Detection

Magnitudes of increase in tRNA detection sensitivity and quantification accuracy with the treated compared with untreated samples.

🧬

Designed for tRNA

Optimized for reverse transcribing tRNAs efficiently, overcoming the barriers that hinder traditional cDNA synthesis.

🔗

System-Compatible

Recommended companion for the nrStar™ tRNA PCR Array and rtStar™ Pre-designed tRNA Primer Sets.

📦

Ready-to-Use Format

Convenient 12-reaction format for direct use in tRNA qPCR workflows.

Background

Why tRNA pretreatment is essential

tRNAs are heavily modified small RNAs: extensive internal and terminal post-transcriptional modifications block reverse transcription and distort readout. These modification barriers prevent regular cDNA synthesis methods from efficiently reverse transcribing tRNAs, causing biased and inaccurate detection.

The rtStar™ tRNA Pretreatment & First-Strand cDNA Synthesis Kit removes internal modifications of tRNAs before first-strand cDNA synthesis, enabling efficient reverse transcription. When applied with the nrStar™ tRNA PCR Array and rtStar™ Pre-designed tRNA Primer Sets, it delivers magnitudes of increase in tRNA detection sensitivity and quantification accuracy.

Kit Workflow

From total RNA to tRNA cDNA in three steps

1

RNA Input

Purified total RNA containing small RNAs is used as input, with quality and quantity assessment.

2

tRNA Pretreatment

Internal modifications of tRNAs are efficiently removed by the pretreatment step, clearing the barriers that block reverse transcriptase.

3

First-Strand cDNA Synthesis

Pretreated tRNAs are reverse transcribed into first-strand cDNA, ready for qPCR with the nrStar™ tRNA PCR Array or rtStar™ Pre-designed tRNA Primer Sets.

Bioinformatics & Deliverables

Product specifications and documentation

The rtStar™ tRNA Pretreatment & First-Strand cDNA Synthesis Kit is a complete, ready-to-use system for efficient tRNA reverse transcription. It is optimized for use with the nrStar™ tRNA PCR Array and rtStar™ Pre-designed tRNA Primer Sets.

Standard Deliverables

Research Applications

Where the rtStar™ kit is used

tRNA qPCR Arrays

Recommended companion for the nrStar™ tRNA PCR Array (Cat# AS-NR-001) for accurate isoacceptor and isodecoder profiling.

Pre-designed Primer Sets

Pairs with rtStar™ Pre-designed tRNA Primer Sets for individual mature tRNA quantification.

tRNA Sequencing

The same pretreatment principle underlies Arraystar tRNA-seq and tRNA modification sequencing services.

Translation Research

Accurate tRNA detection supports studies of codon usage, translation efficiency, and tRNA biology.

Sample Requirements

Recommended RNA input for rtStar™ kit projects

RNA Amount & Quality

  • Total RNA input: > 5 µg per sample is recommended for tRNA qPCR array projects (official Arraystar recommended minimum for the entire experiment in a single attempt, including sample QC).
  • 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: required because the sample is 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 and the sample list.
  • Ship to: Arraystar Inc., 9430 Key West Avenue #128, Rockville, MD 20850, USA. Contact us before shipping.

FAQ

Common questions about the rtStar™ kit

Why is tRNA pretreatment necessary before cDNA synthesis?
tRNAs carry extensive internal post-transcriptional modifications that block reverse transcriptase during cDNA synthesis, causing biased and inaccurate detection. Regular cDNA synthesis methods cannot remove these barriers. The rtStar™ kit efficiently removes internal modifications of tRNAs, ensuring the proceeding of reverse transcriptase for accurate cDNA synthesis.
What improvement can I expect with the treated samples?
The kit delivers magnitudes of increase in tRNA detection sensitivity and quantification accuracy with the treated compared with untreated samples. This improvement applies to both qPCR array profiling and individual tRNA quantification with pre-designed primer sets, because the pretreatment clears the modification barriers that block reverse transcriptase.
Which products is this kit optimized for?
The kit is designed for reverse transcribing tRNAs efficiently and is the recommended companion for the nrStar™ tRNA PCR Array (Cat# AS-NR-001) and rtStar™ Pre-designed tRNA Primer Sets, together with Arraystar SYBR® Green Real-time qPCR Master Mix for optimal performance.
What is the recommended RNA input?
We recommend more than 5 µg of total RNA per sample for tRNA qPCR array projects — 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. DNase treatment is required because the sample is used for qPCR.
How many reactions are included?
The kit is supplied in a 12-reaction format (Catalog No. AS-FS-004), sufficient for standard tRNA pretreatment and first-strand cDNA synthesis experiments. A product manual is included with detailed protocols, and the kit is designed to pair with the nrStar™ tRNA PCR Array and pre-designed primer sets.
Can the kit be used for tRNA sequencing?
The pretreatment principle of removing internal tRNA modifications is the foundation of Arraystar's tRNA sequencing methodologies. For sequencing projects, Arraystar offers end-to-end service options, and the rtStar™ tRNA Pretreatment Kit is also part of the tRF&tiRNA and tRNA sequencing workflows.

Selected Publications

Featured Client Publications Using rtStar™ tRNA Pretreatment

  1. Kalita B, et al. PAX translocations remodel mitochondrial metabolism through altered leucine usage in rhabdomyosarcoma. Cell, 2025. PMID: 40185100
  2. El-Hachem N, et al. Valine aminoacyl-tRNA synthetase promotes therapy resistance in melanoma. Nature Cell Biology, 2024. PMID: 38849541
  3. Shen Z, et al. tRNA Ini CAT inhibits proliferation and promotes apoptosis of laryngeal squamous cell carcinoma cell. Journal of Clinical Laboratory Analysis, 2021. PMID: 34048096
  4. Gerber A, et al. Gene-Specific Control of tRNA Expression by RNA Polymerase II. Molecular Cell, 2020. PMID: 32298650
  5. Pinkard O, et al. Quantitative tRNA-sequencing uncovers metazoan tissue-specific tRNA regulation. Nature Communications, 2020. PMID: 32796835
  6. Kuang M, et al. tRNA-based prognostic score in predicting survival outcomes of lung adenocarcinomas. International Journal of Cancer, 2019. PMID: 30838640
  7. Li M, et al. DNA damage-induced cell death relies on SLFN11-dependent cleavage of distinct type II tRNAs. Nature Structural & Molecular Biology, 2018. PMID: 30374083

Ready to Achieve Accurate tRNA Detection?

The rtStar™ tRNA Pretreatment & First-Strand cDNA Synthesis Kit removes tRNA modification barriers for efficient, accurate cDNA synthesis — request a quote today.