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Updated: Sep 25, 2025

Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing

Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing

Published on: November 18, 2014

Intramolecular ligation method (iLIME) for pre-miRNA quantification and sequencing.

Minh Ngoc Le¹, Cong Truc Le¹, Tuan Anh Nguyen¹

¹Division of Life Science, The Hong Kong University of Science and Technology, Hong Kong, China.

RNA (New York, N.Y.)

|April 29, 2022

Summary

This summary is machine-generated.

We developed the intramolecular ligation method (iLIME) to specifically quantify and sequence precursor microRNAs (pre-miRNAs). This novel technique accurately measures pre-miRNAs, distinguishing them from pri-miRNAs for better disease research.

Keywords:

microRNA pre-miRNA pre-miRNA quantification pre-miRNA sequencing pri-miRNA

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Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Published on: April 14, 2015

Area of Science:

Molecular Biology
Genetics
Biochemistry

Background:

MicroRNAs (miRNAs) are crucial regulators of gene expression involved in numerous human diseases.
Precursor microRNAs (pre-miRNAs) are essential intermediates in miRNA biogenesis.
Existing quantification methods struggle to differentiate between pre-miRNAs and their parental pri-miRNAs.

Purpose of the Study:

To develop a method for specific quantification and sequencing of pre-miRNAs.
To overcome limitations of current quantitative PCR (qPCR) methods in distinguishing pre-miRNAs from pri-miRNAs.
To provide a tool for enhanced investigation of pre-miRNA roles in biological processes and diseases.

Main Methods:

Development of the intramolecular ligation method (iLIME).
Utilization of T4 RNA ligase 1 to circularize pre-miRNAs.
Design of specific PCR primers for pre-miRNA amplification, excluding pri-miRNAs.
Integration of next-generation sequencing for pre-miRNA end sequencing.

Main Results:

iLIME enables precise quantification of pre-miRNAs.
The method effectively distinguishes pre-miRNAs from pri-miRNAs.
iLIME allows for sequencing of pre-miRNA ends.
Successful application of iLIME for pre-miRNA analysis.

Conclusions:

iLIME is a simple and effective method for specific pre-miRNA quantification and sequencing.
This technique offers significant advantages over current methods for pre-miRNA research.
iLIME holds promise for advancing studies in gene regulation, human diseases, and medical applications.