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Related Concept Videos

DNA Microarrays02:34

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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: May 19, 2026

Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations
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Multiplex cDNA quantification method that facilitates the standardization of gene expression data.

Osamu Gotoh1, Yasufumi Murakami, Akira Suyama

  • 1Department of Life Sciences and Institute of Physics, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan.

Nucleic Acids Research
|March 19, 2011
PubMed
Summary

This study introduces GEP-DEAN, a novel method for measuring absolute gene expression levels using DNA microarrays. This approach overcomes limitations of relative quantification, enabling more reliable transcriptome analysis.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Microarray-based gene expression analysis is crucial for transcriptome studies.
  • Current methods provide only relative gene expression levels, affected by platforms and RNA references.
  • Valid comparisons necessitate standardized platforms, controls, and complex normalization, limiting extensive data analysis.

Purpose of the Study:

  • To develop an effective approach for measuring absolute gene expression levels on common DNA microarrays.
  • To overcome the limitations of relative quantification in microarray data analysis.
  • To enable more reliable and extensive comparisons of gene expression data.

Main Methods:

  • Development of a multiplex cDNA quantification method named GEP-DEAN (Gene expression profiling by DCN-encoding-based analysis).
  • Utilizing common DNA microarrays for absolute quantification.
  • Validation using chemically synthesized DNA strands and mouse liver cDNA samples.

Main Results:

  • Demonstrated successful measurement of absolute cDNA strand amounts.
  • Achieved a sensitivity of 18 zmol.
  • The method operates in a highly multiplexed manner within 7 hours.

Conclusions:

  • GEP-DEAN offers an effective solution for absolute gene expression quantification on DNA microarrays.
  • This method removes limitations associated with relative quantification, facilitating more robust transcriptome analysis.
  • The developed technique shows high sensitivity and efficiency for multiplexed gene expression profiling.