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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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qPCRTag Analysis - A High Throughput, Real Time PCR Assay for Sc2.0 Genotyping
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Highly sensitive genotyping using artificial riboregulator system.

Atsushi Narita1, Kazumasa Ogawa, Shinsuke Sando

  • 1Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto university, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.

Nucleic Acids Symposium Series (2004)
|December 8, 2006
PubMed
Summary

This study introduces a novel riboregulator for sensitive gene detection. The system achieves high sensitivity and single-nucleotide accuracy for detecting target genes using RNA or DNA probes.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Biochemistry

Background:

  • Gene sensing technologies are crucial for molecular diagnostics.
  • Existing methods often require complex sample preparation or lack sensitivity.
  • Riboregulators offer a potential platform for simplified and sensitive molecular detection.

Purpose of the Study:

  • To develop a novel molecular-beacon-type riboregulator for enhanced catalytic gene sensing.
  • To demonstrate the system's capability for detecting target genes using RNA or dsDNA probes.
  • To evaluate the sensitivity and specificity of the developed riboregulator system.

Main Methods:

  • Constructed a riboregulator system incorporating a firefly luciferase reporter gene.
  • Engineered a regulator hairpin domain with a ribosome binding site (RBS)/anti-RBS stem and a target-complementary loop.
  • Utilized unmodified RNA or dsDNA as probes for target gene sensing.
  • Measured gene sensing via chemiluminescence output.

Main Results:

  • Achieved highly sensitive gene detection with a lower limit of < or = 50 fmol of the target.
  • Demonstrated single-nucleotide resolution in target gene identification.
  • Successfully employed both RNA and dsDNA probes for sensing.
  • Validated the catalytic amplification of gene sensing.

Conclusions:

  • The developed molecular-beacon-type riboregulator enables sensitive and specific gene detection.
  • This system offers a promising platform for nucleic acid sensing applications.
  • The demonstrated sensitivity and resolution pave the way for advanced diagnostic tools.