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Real Time RT-PCR

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

PCR

PCR

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Real-time loop-mediated isothermal DNA amplification in compact disc micro-reactors.

Sara Santiago-Felipe¹, Luis A Tortajada-Genaro¹, Javier Carrascosa¹

¹Instituto interuniversitario de Reconocimiento Molecular y Desarrollo Tecnológico (IDM), Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, E46071 Valencia, Spain.

Biosensors & Bioelectronics

|December 31, 2015

Summary

This summary is machine-generated.

This study introduces a compact disc-based device for rapid DNA detection using in-disc loop-mediated isothermal amplification (iD-LAMP). The system offers portable, low-cost, high-throughput analysis with high sensitivity for pathogen and food identification.

Keywords:

Compact disc DNA amplification Isothermal real-time LAMP Micro-reactors

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Rapid PCR Thermocycling using Microscale Thermal Convection

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Rapid PCR Thermocycling using Microscale Thermal Convection

Rapid PCR Thermocycling using Microscale Thermal Convection

Published on: March 5, 2011

Area of Science:

Biotechnology
Analytical Chemistry
Molecular Diagnostics

Background:

Real-time DNA determination is crucial for diagnostics and food safety.
Conventional methods can be time-consuming and require specialized equipment.
Isothermal amplification offers advantages over traditional PCR but requires efficient detection systems.

Purpose of the Study:

To develop an integrated device for real-time targeted DNA determination using micro-reactors on compact discs.
To leverage in-disc loop-mediated isothermal amplification (iD-LAMP) with quantitative optical readout.
To demonstrate the device's utility for detecting pathogens and identifying food components.

Main Methods:

Micro-reactors were fabricated on digital video discs (DVDs).
In-disc loop-mediated isothermal amplification (iD-LAMP) was employed for DNA amplification.
Quantitative optical readout using a disc drive monitored changes in turbidity or colorimetry.

Main Results:

The developed iD-LAMP assay achieved high sensitivity (5 CFU/mL for bacteria, 10 µg/g for meat).
Simultaneous analysis of 96 samples was completed in 15 minutes.
Results from the disc-based device correlated well with conventional LAMP and qPCR methods.

Conclusions:

The compact disc-integrated device provides a portable, cost-effective, and high-throughput platform for rapid DNA analysis.
The system enables low sample and reagent consumption with ease of use.
This technology holds promise for point-of-care diagnostics and food safety applications.