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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...

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Related Experiment Video

Updated: Jul 2, 2025

Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

Published on: September 24, 2015

A highly integrated digital PCR system with on-chip heating for accurate DNA quantitative analysis.

Kang Peng¹, Zhihong Wu¹, Zhongxin Feng²

¹BOE Technology Group Co Ltd., Beijing, 100176, PR China.

Biosensors & Bioelectronics

|February 29, 2024

Summary

A novel self-heating digital polymerase chain reaction (dPCR) chip offers faster, more efficient disease diagnosis. This integrated system improves sample handling and thermal cycling, making sensitive molecular detection more accessible.

Keywords:

BCR-ABL1 mutation detection Heterogeneous modification Mass production process Rapid digital PCR thermocycling

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Area of Science:

Biotechnology
Molecular Diagnostics
Microfluidics

Background:

Digital polymerase chain reaction (dPCR) enables highly sensitive disease diagnosis via single-molecule detection.
Current dPCR systems face limitations including complex sample distribution, external heaters, and slow thermal cycling, hindering efficiency.

Purpose of the Study:

To develop an improved dPCR system with an integrated self-heating chip for enhanced speed and efficiency.
To optimize chip design for better sample partitioning and thermal uniformity.

Main Methods:

Microwell array dPCR system with an integrated self-heating chip.
Hydrodynamic and electrothermal simulations for chip optimization.
Hydrophilic/hydrophobic surface modifications for sample compartmentalization.
Indium tin oxide (ITO) heating layer and a custom temperature control algorithm for rapid, stable self-heating.

Main Results:

Optimized chip design improved microwell partitioning and thermal distribution.
The system demonstrated a dynamic linear range of 10^5 and a detection limit of 10 copies/reaction for EGFR plasmid DNA.
Quantitative detection of BCR-ABL1 mutation gene fragments showed comparable analytical accuracy to a commercial dPCR system.

Conclusions:

The developed self-heating dPCR system significantly enhances efficiency and speed.
Cost-effective chip manufacturing via semiconductor lines promotes widespread adoption and accessibility.
This integrated dPCR platform offers a promising solution for sensitive and efficient molecular diagnostics.