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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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High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR
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High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR

Published on: November 28, 2016

Nanoliter high throughput quantitative PCR.

Tom Morrison1, James Hurley, Javier Garcia

  • 1BioTrove Inc, 12 Gill Street, Suite 4000, Woburn, MA 01810, USA.

Nucleic Acids Research
|September 27, 2006
PubMed
Summary

This study introduces a novel high-throughput method for gene expression analysis, combining real-time PCR (RT-PCR) accuracy with microarray parallelism. This approach significantly enhances RNA quantification efficiency for biological complexity research.

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Last Updated: Jul 19, 2026

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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

Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Accurate RNA quantification is crucial for understanding gene expression patterns and biological complexity.
  • Traditional methods like real-time PCR (RT-PCR) and microarrays have limitations in throughput, accuracy, or sensitivity.
  • Scaling RT-PCR is hindered by cost and logistics, while microarrays suffer from low sensitivity and dynamic range.

Purpose of the Study:

  • To develop a hybrid approach combining RT-PCR accuracy with microarray parallelism for high-throughput gene expression analysis.
  • To overcome the limitations of existing quantitative transcriptional analysis methods.

Main Methods:

  • Development of a microfluidic device containing an array of 3072 real-time polymerase chain reactions (RT-PCRs) in 33 nl volumes.
  • Integration of RT-PCR's precision and dynamic range with microarray's parallelism on a microscope slide-sized platform.

Main Results:

  • Achieved RT-PCR accuracy and precision equivalent to standard 384-well microplate assays.
  • Demonstrated a 64-fold reduction in reaction volume and a 24-fold increase in analytical throughput.
  • Ensured workflow compatibility with standard microplate protocols.

Conclusions:

  • The novel hybrid RT-PCR array offers a significant advancement in high-throughput quantitative transcriptional analysis.
  • This method provides a cost-effective and efficient solution for studying biological complexity through gene expression profiling.
  • The technology enables accurate and precise RNA level measurements across thousands of genes simultaneously.