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Related Experiment Video

Updated: May 15, 2026

Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing
14:15

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

Published on: November 18, 2014

Splinted ligation method to detect small RNAs.

Timothy W Nilsen

    Cold Spring Harbor Protocols
    |January 4, 2013
    PubMed
    Summary
    This summary is machine-generated.

    This study introduces a splinted ligation method for detecting small RNAs, like microRNAs (miRNAs) and piwi-associated RNAs (piRNAs). This technique quantifies specific RNA molecules within complex cellular mixtures.

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    Area of Science:

    • Molecular Biology
    • Genomics
    • Biochemistry

    Background:

    • Cellular RNA populations are highly complex, with diverse sizes and abundances.
    • Accurate detection and quantification of specific small RNA species are crucial for biological research.
    • Existing methods may face challenges in complex RNA mixtures.

    Purpose of the Study:

    • To describe a robust protocol for the detection and quantification of small RNAs.
    • To provide a reliable method for analyzing specific RNA species, including miRNAs and piRNAs.
    • To enable routine analysis of small RNA integrity and abundance.

    Main Methods:

    • The protocol utilizes a splinted ligation approach with T4 DNA ligase.
    • A DNA splint oligonucleotide bridges the RNA of interest and a labeled DNA probe.

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    Last Updated: May 15, 2026

    Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing
    14:15

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

    Published on: November 18, 2014

    Locked Nucleic Acid Flow Cytometry-fluorescence in situ Hybridization (LNA flow-FISH): a Method for Bacterial Small RNA Detection
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  • Ligation covalently joins the RNA and the labeled probe, followed by gel electrophoresis and phosphorimaging for visualization.

    • Main Results:

    • The splinted ligation method allows for the detection of specific small RNA molecules.
    • The method enables the quantification of RNA abundance and assessment of integrity.
    • Visualization via denaturing gel electrophoresis and phosphorimaging confirms successful ligation and detection.

    Conclusions:

    • The described splinted ligation protocol is effective for detecting and quantifying small RNAs.
    • This method is suitable for routine analysis of microRNAs (miRNAs) and piwi-associated RNAs (piRNAs).
    • The technique offers a valuable tool for studying complex RNA mixtures in cellular systems.