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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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Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
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Quantitative PCR based expression analysis on a nanoliter scale using polymer nano-well chips.

Andreas Dahl1, Marc Sultan, Alexander Jung

  • 1Max-Planck-Institute for Molecular Genetics, Department Vertebrate Genomics, Ihnestrasse 73, 14195 Berlin-Dahlem, Germany. dahl@molgen.mpg.de

Biomedical Microdevices
|January 5, 2007
PubMed
Summary

This study introduces a miniaturized platform for quantitative PCR (qPCR) enabling high-throughput gene expression analysis. The microPCR chip offers accurate results in nanoliter volumes, reducing costs and improving efficiency.

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Area of Science:

  • Functional genomics
  • Molecular biology
  • Biotechnology

Background:

  • Gene expression analysis is crucial for understanding biological functions.
  • DNA microarrays offer high throughput but lack the precision of qPCR.
  • Quantitative PCR (qPCR) is the gold standard for validating gene expression data.

Purpose of the Study:

  • To develop a miniaturized, cost-effective platform for qPCR-based gene expression analysis.
  • To combine the sensitivity of qPCR with high data density in a micro-array format.
  • To enable nanoliter-scale liquid PCR assays for enhanced throughput and reduced cost.

Main Methods:

  • Development of an open-well, miniaturized platform using cost-effective polypropylene microreactors (microPCR Chip).
  • Utilizing TaqMan chemistry for qPCR assays in 200 nanoliter volumes.
  • Performing RNA expression analysis on four genes in mouse tissues (brain, liver, kidney).

Main Results:

  • Demonstrated quantification ability and reliability of qPCR in 200 nl volumes down to 5 starting target molecules.
  • Achieved comparable results to standard 10 microliter assays for RNA expression analysis.
  • Validated the sensitivity and quantification capability of the microPCR chip platform.

Conclusions:

  • The microPCR chip is a reliable and sensitive platform for nanoliter-scale qPCR assays.
  • This technology enables high-throughput gene expression analysis with reduced costs.
  • The platform shows promise for advancing functional genomics research through miniaturized qPCR.