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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...
PCR01:32

PCR

Overview

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Related Experiment Video

Updated: Jun 13, 2026

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
10:28

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Published on: April 14, 2015

Different real-time PCR systems yield different gene expression values.

Shan Lu1, Andrew P Smith, Dan Moore

  • 1California Pacific Medical Center Research Institute, San Francisco, CA 94107, USA.

Molecular and Cellular Probes
|April 27, 2010
PubMed
Summary
This summary is machine-generated.

Real-time polymerase chain reaction (PCR) systems show significant differences. Gene expression analysis may vary between platforms like ABI and Roche, impacting research reproducibility.

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Last Updated: Jun 13, 2026

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genomics

Background:

  • Real-time polymerase chain reaction (PCR) systems utilize diverse settings and proprietary reagents.
  • Variability in PCR platforms may influence gene expression quantification.
  • The impact of inter-platform differences on gene expression data is not well-established.

Purpose of the Study:

  • To compare gene expression analysis between two major real-time PCR systems: Life Technologies (ABI7500) and Roche Applied Science (LC480).
  • To investigate the effect of system-specific parameters and reagents on gene detection and quantification.
  • To assess potential discrepancies in gene expression values obtained from different PCR platforms.

Main Methods:

  • Comparative analysis of ABI7500 and LC480 real-time PCR systems.
  • Utilized default settings, proprietary reagents, and varied parameters (ramp rates, magnesium concentrations).
  • Analyzed expression of four genes (IL-8, COX2, ID-1, CXCR2) in a human breast cancer cell line.

Main Results:

  • Two target genes were detected by ABI7500 but not by the Roche LC480 system under default conditions.
  • Optimization of Roche system parameters partially restored detection but yielded significantly lower expression levels compared to ABI.
  • Even with alternative primer pairs for ID-1, expression values on the LC480 remained substantially lower than on the ABI7500.

Conclusions:

  • Significant technical differences exist between ABI7500 and LC480 real-time PCR systems.
  • These platform-specific variations can lead to substantial discrepancies in gene expression data.
  • Researchers must consider potential inter-platform variability when interpreting and reporting gene expression results.