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Related Concept Videos

Southern Blot02:57

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
In-situ Hybridization02:31

In-situ Hybridization

In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
FISH - Fluorescent In-situ Hybridization02:07

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...
Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

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Related Experiment Video

Updated: May 19, 2026

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method
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Detecting miRNAs by liquid hybridization and color development.

Xiangqi Li1, Minjie Ni, Yonglian Zhang

  • 1Shanghai Key Laboratory for Molecular Andrology, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Shanghai 200031, China. lixq@sibs.ac.cn

Methods (San Diego, Calif.)
|August 14, 2012
PubMed
Summary

We developed a new method, liquid hybridization and color development (LHCD), for detecting microRNAs (miRNAs). This simple, visual technique offers sensitive and rapid miRNA detection without expensive equipment.

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Area of Science:

  • Molecular Biology
  • Biochemistry

Background:

  • Polymerase chain reaction (PCR) and Northern blot are common microRNA (miRNA) detection methods.
  • PCR suffers from false positives and primer design issues, while Northern blots are complex and time-consuming.
  • Existing rapid liquid Northern blot methods require specialized equipment for signal detection.

Purpose of the Study:

  • To introduce a novel, simplified protocol for microRNA detection.
  • To provide a sensitive, rapid, and visually detectable method for miRNA analysis.
  • To overcome the limitations of current miRNA detection techniques.

Main Methods:

  • Developed a liquid hybridization and color development (LHCD) assay.
  • Utilized signal amplification via the avidin-biotin complex (ABC) system.
  • Employed visual detection, eliminating the need for specialized instruments.

Main Results:

  • The LHCD protocol achieved sensitive detection of 2.5 fmol of miRNAs.
  • LHCD demonstrated the ability to differentiate between miRNAs with single-nucleotide variations.
  • The method proved to be simple, rapid, and visually based.

Conclusions:

  • LHCD offers a convenient and accessible alternative for microRNA detection.
  • The protocol is easy to learn and suitable for routine miRNA analysis.
  • LHCD provides a cost-effective and efficient solution for miRNA research.