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tRF&tiRNA Research Tools

nrStar™ tRF&tiRNA PCR Array (H/M) — High-Specificity Profiling of tRFs and tiRNAs

nrStar™ Human tRF & tiRNA PCR Array profiles 185 prevalent tRFs or tiRNAs compiled in the databases and newly reported in recent publications. nrStar™ Mouse tRF PCR Array profiles 88 prevalent tRFs selected from the tRFdb database. Applied with the rtStar™ tRF&tiRNA Pretreatment Kit (CAT# AS-FS-005), the PCR Arrays offer a new level of accuracy on tRF&tiRNA detection.

384-well ready-to-run format · adaptor-based specificity · 185 human / 88 mouse targets

Overview

Ready-to-run qPCR panels with precursor discrimination

nrStar™ Human tRF & tiRNA PCR Array profiles 185 prevalent tRFs or tiRNAs compiled in the databases and newly reported in recent publications. nrStar™ Mouse tRF PCR Array profiles 88 prevalent tRFs selected from the tRFdb database.

In order to distinguish tRF&tiRNA from its precursor and achieve high specificity, the rtStar™ First-Strand cDNA Synthesis Kit (3′ and 5′ adaptor) (CAT# AS-FS-003) is required to be used with the nrStar™ tRF&tiRNA PCR Array for tRF&tiRNA detection.

What is the nrStar™ tRF&tiRNA PCR Array?

The nrStar™ tRF&tiRNA PCR Array is a 384-well, ready-to-run qPCR panel that profiles 185 prevalent human tRFs/tiRNAs (or 88 mouse tRFs). By introducing 3′ and 5′ adaptors, the assay discriminates tRFs/tiRNAs from their tRNA or pre-tRNA precursors for specific detection.

Service at a Glance

Standard catalog products — human and mouse versions available

Product NameCatalog No.SizePrice
nrStar™ Human tRF&tiRNA PCR ArrayAS-NR-002-1384-well (2*192) / plate
nrStar™ Human tRF&tiRNA PCR Array (Roche Light Cycler 480)AS-NR-002-1-R384-well (2*192) / plate
nrStar™ Mouse tRF PCR ArrayAS-NR-002M-1384-well (2*192) / plate

Benefits

Why researchers choose the nrStar™ tRF&tiRNA PCR Array

🎯

Focused Content

The prevalent Human tRFs&tiRNAs/Mouse tRFs having the highest biological potentials are profiled.

Rigorous Design

All primer pairs are meticulously designed, optimized, and validated.

Convenient Format

Easy-to-use, ready-to-run, standard qPCR plate format for direct sample application. No sample pre-amplification is needed.

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Precursor Discrimination

Adaptor-based assay effectively discriminates tRF&tiRNA from their precursors and other small RNAs.

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New Level of Accuracy

Applied with the rtStar™ tRF&tiRNA Pretreatment Kit (CAT# AS-FS-005) for a new level of accuracy on tRF&tiRNA detection.

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System-Compatible

Requires the rtStar™ First-Strand cDNA Synthesis Kit (3′ and 5′ adaptor) (CAT# AS-FS-003) for tRF&tiRNA detection.

Background

Why tRF&tiRNA detection needs adaptor-based discrimination

tRFs & tiRNAs perform many biological functions: they act as microRNAs in RNA interference; regulate target mRNA stability; assemble stress granules in response to cellular stress; modulate apoptosis; and pass on as epigenetic factors in intergenerational inheritance [1] (Fig. 1). The composition and abundance of tRFs&tiRNAs are highly dependent on the cell types and are associated with many disease conditions. The highly enriched presence in biofluids, often much more so than microRNAs, makes them excellent biomolecules for biomarkers [2-5].

tRF&tiRNAs are generated from tRNA or pre-tRNA through precise biogenesis processes. It is difficult to distinguish tRF&tiRNA from its precursor (tRNA or pre-tRNA) by conventional qPCR for their sharing of the same sequence. Moreover, the length of tRFs&tiRNAs is 16~50 nt, too short to perform the qPCR process.

Arraystar exploits a smart system by introducing two artificial sequences to the 3′ and 5′ ends of tRF&tiRNA, a famous and validated technology used in small RNA library preparation for sequencing. Corresponding forward and reverse primers targeting the 5′ and 3′ junction effectively discriminate tRF&tiRNA from its precursors and other small RNA, thus getting a reliable and accurate detection of tRF&tiRNA expression (Fig. 2).

Figure 1. tRF & tiRNA functions and associated diseases.
Figure 1. tRF & tiRNA functions and associated diseases.
Figure 2. By introducing 5′ and 3′ adaptors, nrStar™ tRF&tiRNA PCR panel detects the specific tRF&tiRNAs with no amplification of tRNAs or pre-tRNAs.
Figure 2. By introducing 5′ and 3′ adaptors, nrStar™ tRF&tiRNA PCR panel detects the specific tRF&tiRNAs with no amplification of tRNAs or pre-tRNAs.

Background References

  1. Anderson P, Ivanov P. tRNA fragments in human health and disease. FEBS Letters, 2014. PMID: 25220675
  2. Telonis AG, et al. Dissecting tRNA-derived fragment complexities using personalized transcriptomes reveals novel fragment classes and unexpected dependencies. Oncotarget, 2015. PMID: 26325506
  3. Olvedy M, et al. A comprehensive repertoire of tRNA-derived fragments in prostate cancer. Oncotarget, 2016. PMID: 27015120
  4. Schageman J, et al. The complete exosome workflow solution: from isolation to characterization of RNA cargo. BioMed Research International, 2013. PMID: 24205503
  5. Dhahbi JM, et al. 5′ tRNA halves are present as abundant complexes in serum, concentrated in blood cells, and modulated by aging and calorie restriction. BMC Genomics, 2013. PMID: 23638709

tRF&tiRNA PCR Array Workflow

From RNA to tRF&tiRNA expression profile

1

RNA Preparation

Total RNA is purified with small-RNA-retaining methods, followed by quality and quantity assessment.

2

rtStar™ Pretreatment

The rtStar™ tRF&tiRNA Pretreatment Kit (CAT# AS-FS-005) removes modifications in tRF & tiRNA before cDNA library construction for qPCR.

3

cDNA Synthesis

The rtStar™ First-Strand cDNA Synthesis Kit (3′ and 5′ adaptor) (CAT# AS-FS-003) ligates adaptors and synthesizes cDNA, enabling precursor discrimination.

4

qPCR Setup & Run

cDNA is applied directly to the ready-to-run 384-well plate with SYBR® Green Master Mix on a compatible instrument.

5

Data Analysis

The analysis tool computes tRF&tiRNA expression with the tRF&tiRNA database annotations.

Figure 1. tRF&tiRNA qPCR array workflow.
Figure 1. tRF&tiRNA qPCR array workflow.

Bioinformatics & Deliverables

Product specifications, database, and tools

The nrStar™ tRF&tiRNA PCR Array System is designed to conveniently and quickly profile the tRFs & tiRNAs with high specificity, composed of two products: the rtStar™ First-Strand cDNA Synthesis Kit (3′ and 5′ adaptor) for adaptor ligation and cDNA synthesis, and the nrStar™ tRF&tiRNA PCR Array for tRF&tiRNA detection.

Standard Deliverables

Compatible qPCR Instruments (384-Well Format Block)

Research Applications

tRF&tiRNA research applications

Biomarker Discovery

The highly enriched presence of tRFs&tiRNAs in biofluids, often much more so than microRNAs, makes them excellent biomolecules for biomarkers.

RNA Interference

tRFs act as microRNAs in RNA interference, participating in small RNA-mediated gene silencing.

mRNA Stability Regulation

tRFs bind protein factors to regulate target mRNA stability.

Apoptosis & Stress Response

tRFs modulate apoptosis and assemble stress granules in response to cellular stress.

Intergenerational Inheritance

tRFs pass on as epigenetic factors in intergenerational inheritance.

Sample Requirements

Recommended RNA input for nrStar™ tRF&tiRNA PCR Array projects

RNA Amount & Quality

  • Total RNA input: > 5 µg per sample for the nrStar™ tRF&tiRNA PCR Array (AS-NR-002) — the official recommended minimum for the entire experiment in a single attempt, including sample QC. Supply twice the recommended minimum to avoid project delays.
  • Purification: TRIzol / RNA precipitation or an RNA isolation kit. Because tRF&tiRNA are < 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 nrStar™ tRF&tiRNA PCR Array

What does the nrStar™ tRF&tiRNA PCR Array profile?
The Human tRF & tiRNA PCR Array profiles 185 prevalent tRFs or tiRNAs compiled in the databases and newly reported in recent publications; the Mouse tRF PCR Array profiles 88 prevalent tRFs selected from the tRFdb database. All target tRFs/tiRNAs have the highest biological potentials.
Why is conventional qPCR inadequate for tRF&tiRNA detection?
tRF&tiRNAs are generated from tRNA or pre-tRNA through precise biogenesis and share the same sequence as their precursors, so conventional qPCR cannot distinguish them. Moreover, their length of 16~50 nt is too short to perform the qPCR process directly, and their modified termini and internal modifications block standard cDNA synthesis.
How does the array discriminate tRFs&tiRNAs from their precursors?
Arraystar introduces two artificial sequences to the 3′ and 5′ ends of tRF&tiRNA, a validated technology used in small RNA library preparation for sequencing. Forward and reverse primers targeting the 5′ and 3′ junction effectively discriminate tRF&tiRNA from precursors and other small RNAs, giving reliable detection.
Which additional products are required?
The rtStar™ tRF&tiRNA Pretreatment Kit (CAT# AS-FS-005) removes modifications in tRF & tiRNA before cDNA library construction for a new level of accuracy, and the rtStar™ First-Strand cDNA Synthesis Kit (3′ and 5′ adaptor) (CAT# AS-FS-003) is required for tRF&tiRNA detection with the array.
What is the recommended RNA input?
We recommend more than 5 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. DNase treatment is required because the sample is used for qPCR.
Which qPCR instruments are compatible?
The array is compatible with instruments equipped with a 384-well format block, including ABI ViiA™ 7, ABI 7500 & 7500 FAST, ABI 7900HT, ABI QuantStudio™ 5/6 Flex/7 Flex/12K Flex, Bio-Rad CFX384, Bio-Rad iCycler & iQ, Eppendorf Realplex, and Roche Light Cycler 480.

Selected Publications

Featured Client Publications in tRF&tiRNA Research

  1. Zhao Y, et al. Differentially expressed tRNA-derived fragments and their roles in primary cardiomyocytes stimulated by high glucose. Frontiers in Endocrinology, 2023. PMID: 36714586
  2. Gornalusse G, et al. Men who inject opioids exhibit altered tRNA-Gly-GCC isoforms in semen. Molecular Human Reproduction, 2023. PMID: 36661332
  3. Xi J, et al. Expression and Diagnostic Value of tRNA-Derived Fragments Secreted by Extracellular Vesicles in Hypopharyngeal Carcinoma. OncoTargets and Therapy, 2021. PMID: 34285510
  4. Kong XY, et al. Deletion of Endonuclease V suppresses chemically induced hepatocellular carcinoma. Nucleic Acids Research, 2020. PMID: 32083667
  5. Zhang F, et al. A 3′-tRNA-derived fragment enhances cell proliferation, migration and invasion in gastric cancer by targeting FBXO47. Archives of Biochemistry and Biophysics, 2020. PMID: 32592804
  6. Cooke WR, et al. Maternal circulating syncytiotrophoblast-derived extracellular vesicles contain biologically active 5′ tRNA halves. Biochemical and Biophysical Research Communications, 2019. PMID: 31405560

Ready to Profile tRFs and tiRNAs with High Specificity?

The nrStar™ tRF&tiRNA PCR Array delivers precursor-discriminated tRF/tiRNA expression profiles in hours — request a quote or ask about the target list.