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Related Concept Videos

PCR01:32

PCR

Overview
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: Jul 11, 2026

An Allele-specific Gene Expression Assay to Test the Functional Basis of Genetic Associations
10:17

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Published on: November 3, 2010

Comparative analysis for expressed genes by polymerase chain reaction using module-shuffling primers.

C Uematsu1, J Nishida, K Okano

  • 1Biosystems Research Department, Central Research Laboratory, Hitachi Ltd.

Nucleic Acids Research. Supplement (2001)
|July 3, 2003
PubMed
Summary

A new method uses competitive PCR and DNA sequencing for gene expression analysis. This technique accurately compares gene amounts, detecting differences as small as 30% in yeast.

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Last Updated: Jul 11, 2026

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Comparative gene expression analysis is crucial for understanding cellular processes.
  • Existing methods may have limitations in sensitivity or throughput.

Purpose of the Study:

  • To develop a novel, sensitive method for comparative gene expression analysis.
  • To enable accurate quantification of gene expression levels from different biological samples.

Main Methods:

  • Development of competitive Polymerase Chain Reaction (PCR) amplification.
  • Utilizing module-shuffling primers for simultaneous amplification.
  • Employing gel electrophoresis in a fluorescent DNA sequencer for analysis.
  • Tagged-cDNA restriction fragments amplified in a single tube.

Main Results:

  • The method achieves consistent amplification efficiency across different targets.
  • It can detect variations in gene expression levels as low as 30%.
  • Successful application demonstrated in comparative gene expression analysis of yeast.

Conclusions:

  • The developed method offers a reliable approach for quantitative gene expression studies.
  • It provides high sensitivity for detecting subtle changes in gene expression.
  • This technique is valuable for research in yeast and potentially other organisms.