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Southern Blot

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...

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Related Experiment Video

Updated: Apr 20, 2026

Visual Detection of Multiple Nucleic Acids in a Capillary Array

Visual Detection of Multiple Nucleic Acids in a Capillary Array

Published on: November 15, 2017

Fast and highly specific DNA-based multiplex detection on a solid support.

Ivan Barišić¹, Verena Kamleithner, Silvia Schönthaler

¹Molecular Diagnostics, AIT Austrian Institute of Technology, Muthgasse 11, 1190, Vienna, Austria, ivan.barisic@ait.ac.at.

Applied Microbiology and Biotechnology

|December 5, 2014

Summary

This study introduces a novel ligation-based DNA probe (LNC) for rapid bacterial pathogen detection. The method achieves high specificity and sensitivity in multiplex assays, enabling faster infectious disease diagnostics.

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Multiplex PCR and Reverse Line Blot Hybridization Assay mPCR/RLB

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Visual Detection of Multiple Nucleic Acids in a Capillary Array

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Multiplex PCR and Reverse Line Blot Hybridization Assay mPCR/RLB

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Area of Science:

Molecular Biology
Microbiology
Genetics

Background:

Accurate and rapid DNA-based diagnostics are crucial for identifying infectious diseases.
Characterizing over 1000 genetic targets, including antibiotic resistance genes and virulence factors, is vital for effective patient treatment.
Existing multiplex detection methods often lack the speed and sensitivity required for timely clinical decisions.

Purpose of the Study:

To develop a novel ligation-based DNA probe (LNC) concept for enhanced multiplex detection of bacterial pathogens.
To combine LNC probes with microarray technology for a fast and specific diagnostic assay.
To evaluate the sensitivity and specificity of the LNC probe method compared to conventional approaches.

Main Methods:

Development of a novel linear chain (LNC) DNA probe design.
Integration of LNC probes with microarray technology for multiplex detection.
Direct use of genomic DNA as a template in ligation reactions without pre-amplification.

Main Results:

The LNC probes accurately identified all tested bacterial species within 1 hour using a 25-multiplex reaction targeting the 16S rRNA gene.
The method detected as few as 10(7) cells directly from genomic DNA, demonstrating high sensitivity without pre-amplification.
Specificity was confirmed by correctly characterizing single nucleotide polymorphisms (SNPs) without false positive signals.
LNC3 probes exhibited a sensitivity at least 10 times higher than conventional microarray probes.

Conclusions:

A fast, simple, highly specific, and sensitive multiplex DNA detection method using LNC probes has been successfully developed.
This novel approach significantly improves upon conventional microarray probe sensitivity and speed.
The LNC probe technology is adaptable for a wide range of diagnostic applications, particularly in infectious disease characterization.