rtStar™ tRF&tiRNA Pretreatment & First-Strand cDNA Synthesis Kit

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rtStar™ tRF&tiRNA Pretreatment Kit prepares tRF & tiRNA samples before the cDNA synthesis used for qPCR. The kit generates terminal 3’-OH and 5’-P ends for adapter ligation by removing 3’-cP and phosphorylation of 5’-OH often present in tRF and tiRNA fragments. Internal m1A, m1G, and m3C are demethylated for efficient cDNA reverse transcription.

rtStar™ tRF&tiRNA First-Strand cDNA Synthesis Kit converts the pretreated tRF and tiRNA fragments into cDNA for qPCR reaction. The method sequentially ligates the 3’-Adaptor to the 3’-ends of the RNAs and the 5’-Adaptor to the 5’-ends of the RNAs.

Benefits

• Efficient removal of internal modifications to ensure the proceeding of reverse transcriptase during cDNA synthesis, whereas the regular cDNA synthesis method doesn’t have. 
• Efficient removal of terminal modifications to ensure terminal 3’-OH and 5’-P ends suitable for the subsequent adapter ligation.
• Capable of distinguishing tRFs & tiRNAs from their precursors.
• Magnitudes of increase in tRF&tiRNA detection sensitivity, quantification accuracy, and assay discriminating power with the treated compared with untreated samples.

Product NameCatalog NoSizePrice
rtStar™ tRF&tiRNA Pretreatment Kit AS-FS-005 12 reactions
$205.00
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rtStar™ tRF&tiRNA First-Strand cDNA Synthesis Kit AS-FS-003 6 reactions
$261.00
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rtStar™ tRF&tiRNA First-Strand cDNA Synthesis Kit AS-FS-003-02 12 reactions
$419.00
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tRFs and tiRNAs, generated through precise biogenesis processes from tRNA, perform many biological functions as small noncoding RNAs and are associated with many diseases and conditions. However, the characteristics of tRFs and tiRNAs pose special challenges for their expression profiling. 

Varying with their particular sources and endoribonuclease cleavages, tRFs and tiRNAs contain many distinct internal modifications and modified termini. While some tRFs and tiRNAs may inherit methylation modifications (such as m1A) and aminoacylated termini from their source tRNAs, tiRNAs, generated by angiogenin, often have 5’-OH and 3’-cyclic phosphate (cP) modifications occurred at the cleavage site. Many current standard cDNA library construction methods, particularly those that target small noncoding RNAs, include adaptor ligation steps in which 5’- and/or 3’- oligonucleotide adaptors are ligated to the 5’ and 3’ ends of the RNA molecule. Because 5’-P and 3-OH at the ends of the RNA substrate are required for adaptor ligation reactions, standard cDNA library construction methods are inadequate for tRF and tiRNA. Importantly, post-transcriptional modifications within the tRF and tiRNA sequence, such as m1A and m3C, also greatly interfere with reverse transcription for cDNA synthesis (Figure 1). In order to efficiently and accurately analyze tRF and tiRNA by qPCR methods, these obstacles must be removed.

tRF_tiRNA_Pretreatment2

Figure 1. Common modifications on tRF&tiRNA interfere with adaptor ligation and cDNA synthesis, which greatly reduce the performance of subsequent cDNA library preparation of qPCR.

 

Moreover, 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. 


In order to achieve high-fidelity and accurate quantification of tRFs&tiRNAs, Arraystar exploit a smart system, composed of two products: rtStar™ tRF&tiRNA Pretreatment Kit and rtStar™ tRF&tiRNA First-Strand cDNA Synthesis Kit. The Pretreatment Kit is designed to remove internal and terminal modifications. By introducing two artificial sequences to the 3’and 5’end of tRF&tiRNA, the First-Strand cDNA Synthesis Kit can effectively discriminate tRF&tiRNA from its precursors and other small RNAs. 

 

rtStar™ tRF&tiRNA Pretreatment Kit (AS-FS-005)

A specific tRNA half, 5’tiRNA-His-GTG, responds to hypoxia via the HIF1a/ANG axis and promotes colorectal cancer progression by regulating LATS2. Tao E W, et al. Journal of Experimental & Clinical Cancer Research, 2021

Clinical diagnostic values of transfer RNA-derived fragment tRF-19-3L7L73JD and its effects on the growth of gastric cancer cells. Shen Y,et al. Journal of Cancer, 2021

Processing by RNase 1 forms tRNA halves and distinct Y RNA fragments in the extracellular environment. Nechooshtan G,et al. Nucleic Acids Research, 2020

Global identification and characterization of tRNA-derived RNA fragment landscapes across human cancers. Sun X, et al. NAR Cancer, 2020

tRNA-Derived Fragments in Podocytes with Adriamycin-Induced Injury Reveal the Potential Mechanism of Idiopathic Nephrotic Syndrome. Li S, et al. BioMed Research International, 2020

Using tRNA halves as novel biomarkers for the diagnosis of gastric cancer. Zhu L, et al. Cancer Biomarkers, 2019

A tRNA fragment, 5'-tiRNAVal, suppresses the Wnt/ß-Catenin signaling pathway by targeting FZD3 in breast cancer. Mo D, et al. Cancer letters, 2019

Profile analysis reveals transfer RNA fragments involved in mesangial cells proliferation. Lu X, et al. Biochemical and Biophysical Research Communications, 2019

Expression analysis of transfer RNA-derived fragments in the blood of patients with moyamoya disease: A preliminary study. Wang C, et al. Molecular medicine reports, 2019

Systematic Analysis of tRNA-Derived Small RNAs Reveals Novel Potential Therapeutic Targets of Traditional Chinese Medicine (Buyang-Huanwu-Decoction) on Intracerebral Hemorrhage. Li P, et al. International Journal of Biological Sciences, 2019

Pancreatic ß-cell tRNA hypomethylation and fragmentation link TRMT10A deficiency with diabetes. Cosentino C, et al. Nucleic Acids Research, 2018

tRNA-Derived Fragments as Novel Predictive Biomarkers for Trastuzumab-Resistant Breast Cancer. Sun C, et al. Cell Physiol Biochem, 2018

 

rtStar™ tRF&tiRNA First-Strand cDNA Synthesis Kit (AS-FS-003/AS-FS-003-02)

A specific tRNA half, 5’tiRNA-His-GTG, responds to hypoxia via the HIF1a/ANG axis and promotes colorectal cancer progression by regulating LATS2. Tao E W, et al. Journal of Experimental & Clinical Cancer Research, 2021

Expression analysis of transfer RNA-derived fragments in the blood of patients with moyamoya disease: A preliminary study. Wang C, et al. Molecular medicine reports, 2019