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PCR

PCR

Next-generation Sequencing

Next-generation Sequencing

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.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....

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Related Experiment Video

Updated: Oct 13, 2025

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Published on: August 3, 2018

Multiplex PCR Design for Scalable Resequencing.

Darren Korbie¹, Matt Trau^2,3

¹Centre for Personalised Nanomedicine, Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD, Australia. d.korbie@uq.edu.au.

Methods in Molecular Biology (Clifton, N.J.)

|November 13, 2021

Summary

This summary is machine-generated.

Multiplex PCR enables simultaneous analysis of multiple genomic regions from diverse DNA sources, offering a cost-effective and scalable solution for genetic research. This method details library preparation for various sequencing platforms.

Keywords:

Bisulfite DNA DNA methylation Multiplex PCR NGS PCR Resequencing SNP screening Sequencing

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Last Updated: Oct 13, 2025

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Area of Science:

Molecular Biology
Genomics
Next-Generation Sequencing (NGS)

Background:

Conventional PCR typically involves a single assay per reaction, limiting throughput and scalability.
Multiplex PCR, analyzing multiple genomic regions simultaneously, is increasingly adopted for efficiency.
Existing multiplex PCR methods require adaptable protocols for diverse DNA sources and sequencing platforms.

Purpose of the Study:

To provide a detailed protocol for designing, screening, and preparing multiplex PCR amplicon libraries.
To support library preparation for major sequencing platforms: Illumina, Ion Torrent, and Oxford Nanopore.
To offer a flexible and adaptable multiplex PCR method for various research needs and buffer systems.

Main Methods:

Design and screening of multiplex PCR assays for multiple genomic targets.
Preparation of multiplex amplicon libraries suitable for high-throughput sequencing.
Adaptation of the protocol for diverse DNA inputs (e.g., human, plant, FFPE, bisulfite-converted DNA).

Main Results:

Successful design and preparation of multiplex amplicon libraries.
Demonstrated compatibility with Illumina, Ion Torrent, and Oxford Nanopore sequencing platforms.
Flexibility in custom panel design, ranging from a few to hundreds of amplicons.

Conclusions:

The described multiplex PCR method is a versatile, scalable, and cost-effective solution for multi-region genomic analysis.
The protocol's adaptability ensures broad applicability across different DNA types and sequencing technologies.
This approach facilitates routine application of multiplex PCR in various research and clinical settings.