tiRNA & tRF Sequencing Service

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tiRNA & tRF Sequencing Service

IntegrateRNA has developed a high-throughput RNA sequencing platform, to investigate the biological functions of small noncoding RNAs of 16-45nt length, especially for tiRNAs and tRFs. We provide a comprehensive solution that covers total RNA preparation, tiRNA & tRF isolation and purification, library construction, sequencing and data analysis. Based on advanced NGS sequencing platform and data analysis pipeline, we assist in novel tiRNA & tRF discovery, mutation characterization, and expression profiling analysis of tiRNAs & tRFs.

tiRNAs (tRNA halves) and tRFs (tRNA-derived fragments) are a recent class of novel small RNAs derived from tRNA, ranging in length from 16 to 50 nt. tiRNAs & tRFs have been suggested to play roles in cell proliferation, priming of viral reverse transcriptases, regulation of gene expression, translation regulation, modulation of the DNA damage response, tumor suppression, and neurodegeneration. In addition, the composition and quantity of tRF & tiRNA are highly dependent on specific cell types and pathological conditions, making them an excellent biomarker with therapeutic potential. tiRNA & tRF sequencing is used to discover novel tiRNAs & tRFs, examine the differential expression of tiRNA & tRF and to characterize variations with single-base resolution.

Workflow of tiRNA & tRF Sequencing

tiRNA & tRF Sequencing Service

As shown, obtaining a premium RNA sequencing library begins with the isolation of tiRNA & tRF RNAs by size fractionation using gel electrophoresis selection or silica spin columns from total RNA. Following RNA adapter ligation using a 5’ adenylated DNA adapter with a blocked 3’end, small RNAs are reverse transcribed, amplified by PCR and sequenced. In order to maximize tiRNA & tRF sequence data quality, the PCR products are selected by gel purification.

Sample Requirements

  • Cell ≥ 2 x 106
  • Tissue ≥ 200mg
  • Whole blood ≥3 ml
  • Total RNA ≥ 10ug (OD260/280 ≥ 1.8; OD260/230≥2.0; RIN≥6.5; 28S:18S≥1.0)

Sequencing Parameters

  • Read length: single-end 50
  • Recommended sequencing depth: ≥ 10 million read pair per sample

Bioinformatics Analysis

  • Summary of length distribution
  • Identification and characterization of tiRNA & tRF
  • Classification and annotation of tiRNA & tRF
  • Quantification and differential expression analysis
  • Functional enrichment analysis

Benefits and Features

  • Performance optimized tiRNAs & tRFs sequencing method with rigorous QC process.
  • Precise annotation and classification system based on tRNA topology and statistical significance.
  • Comprehensive tiRNAs & tRFs collection from all databases.
  • Data visualization with publication quality graphics.
  • Super cost-effective - simultaneous detection and analysis of tiRNA and tRF, as well as miRNA.
  • One-stop service - customers only need to provide cells, tissues, body fluids or total RNA. IntegrateRNA completes the entire service process from sample preparation, library preparation, sequencing to data analysis.

IntegrateRNA utilizes the Illumina HiSeq platforms to sequence tiRNA & tRF. We have flexible strategies for tiRNA (29-50nt) and tRF (16-28nt) discovery and profiling. Our highly experienced expert team executes quality management, following every procedure to ensure confident and unbiased results. If you have any additional requirements or questions, please do not hesitate to contact us at or 1-631-626-9181. Our technical supports are pleased to solve your problems.

Reference:

  1. Xie, Y. et al (2020). Action mechanisms and research methods of trna-derived small rnas. Signal Transduction and Targeted Therapy, 5(1), 109.
For research use only. Not intended for any clinical use.
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