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Next-generation Sequencing

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The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
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Related Experiment Video

Updated: May 23, 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

Multiplex loop-mediated isothermal amplification detection by sequence-based barcodes coupled with nicking

Chao Liang¹, Yanan Chu, Sijia Cheng

¹Department of Pharmacology, Jinling Hospital, Nanjing University School of Medicine, Nanjing, Jiangsu, PR China.

Analytical Chemistry

|March 28, 2012

Summary

Multiplex loop-mediated isothermal amplification (LAMP) now allows simultaneous detection of multiple pathogens. This novel barcoding method combined with nicking endonuclease-mediated pyrosequencing simplifies complex amplicon analysis for reliable identification.

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Last Updated: May 23, 2026

Visual Detection of Multiple Nucleic Acids in a Capillary Array

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Multiplex Detection of Bacteria in Complex Clinical and Environmental Samples using Oligonucleotide-coupled Fluorescent Microspheres

Multiplex Detection of Bacteria in Complex Clinical and Environmental Samples using Oligonucleotide-coupled Fluorescent Microspheres

Published on: October 23, 2011

Area of Science:

Molecular Biology
Nucleic Acid Amplification
Diagnostic Assays

Background:

Loop-mediated isothermal amplification (LAMP) offers high specificity for DNA replication.
Multiplex detection using LAMP is challenging due to complex amplicon structures.

Purpose of the Study:

To develop a method for simultaneous detection of multiple LAMP products.
To overcome the limitations of multiplex LAMP detection.

Main Methods:

Designed target-specific barcodes and tagged LAMP amplicons using FIP primers.
Introduced nicking endonuclease (NEase) recognition sequences into FIP primers.
Decoded barcoded amplicons using NEase-mediated pyrosequencing starting at the nicked 3' end.

Main Results:

Successfully detected four pathogens (Hepatitis B Virus, Hepatitis C Virus, Human Immunodeficiency Virus, Treponema pallidum) in a single 4-plex LAMP reaction.
Barcodes were designed with reporter and stuffer bases for single-peak pyrogram decoding.
Nicking reaction occurred near barcode regions, enabling precise pyrosequencing initiation.

Conclusions:

Barcoded LAMP coupled with NEase-mediated pyrosequencing provides a simple, rapid, and reliable method for multiplex target identification.
This approach is effective for detecting multiple blood-borne pathogens simultaneously.
The method enhances the utility of LAMP for complex diagnostic applications.