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Related Concept Videos

Real Time RT-PCR02:57

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: Jun 21, 2026

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
07:27

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

Published on: August 3, 2011

[Development, optimization and application of the expression analysis platform based on multiplex quantitative RT-PCR

Qin-Xi Wang1, Kai Li, Yu-Xun Zhou

  • 1Institute of Biological Science & Biotechnology, Donghua University, Shanghai 201620, China. wqx457@163.com

Yi Chuan = Hereditas
|July 10, 2009
PubMed
Summary
This summary is machine-generated.

A novel multiplex quantitative RT-PCR method was developed for accurate gene expression profiling. This technique identified PHF6 as a differentially expressed gene in mouse strains, aiding puberty onset research.

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Area of Science:

  • Molecular Biology
  • Genetics

Context:

  • Gene expression profiling is crucial for understanding biological processes.
  • Existing methods like cDNA chips and real-time quantitative PCR have limitations in accuracy and throughput.
  • Multiplex quantitative RT-PCR offers a complementary approach to overcome these challenges.

Purpose:

  • To establish a multiplex quantitative RT-PCR technology using a universal fluorescent primer.
  • To optimize the method for accurate and reliable gene expression quantification.
  • To identify differentially expressed genes related to mouse puberty onset.

Summary:

  • A chimeric-primer-induced-universal-primer amplification method was developed for multiplex quantitative RT-PCR.
  • The technique ensures target genes are amplified in a constant ratio, offering cost-effectiveness and moderate throughput.
  • Sensitivity was determined to be 10^2 copies, and the optimal primer concentration ratio was found to be 1:1.
  • Touchdown PCR with universal primers enhanced the amplification of low-abundance genes.

Impact:

  • The developed method improves the overall process of gene expression profiling analysis.
  • It provides a reliable and accurate alternative for gene expression quantification.
  • The study identified PHF6 as a differentially expressed gene in specific mouse strains, providing a candidate for further functional analysis in puberty onset research.