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Related Concept Videos

RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer.
Since the...
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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.
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Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp
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RT-isoPCR: nested, high multiplex mRNA amplification.

Martin Jensen Søe1, Peter Warthoe

  • 1Atonomics, Vestre Teglgade 10, DK-2450 Copenhagen SV, Denmark. martin.j.soe@gmail.com pw@atonomics.com.

The Analyst
|August 22, 2013
PubMed
Summary
This summary is machine-generated.

RT-isoPCR enables multiplex amplification of mRNA targets. This method achieves sensitive and specific detection of 24 targets without introducing bias, advancing molecular diagnostics.

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Last Updated: May 8, 2026

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Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations
10:23

Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations

Published on: January 19, 2017

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genomics

Background:

  • Multiplex amplification of mRNA targets is crucial for gene expression analysis.
  • Existing methods can face challenges with sensitivity, specificity, and bias.
  • Accurate detection of multiple mRNA targets requires robust amplification techniques.

Purpose of the Study:

  • To introduce and validate RT-isoPCR, a novel method for multiplex mRNA amplification and detection.
  • To assess the sensitivity, specificity, and bias of RT-isoPCR for detecting numerous mRNA targets simultaneously.
  • To demonstrate the utility of RT-isoPCR in molecular diagnostics and gene expression profiling.

Main Methods:

  • Utilized a two-stage approach: multiplex reverse transcription polymerase chain reaction (RT-PCR) followed by isothermal amplification.
  • Developed a system for the individual target loci detection after the initial multiplex amplification.
  • Applied the method to detect a panel of 24 distinct mRNA targets.

Main Results:

  • Achieved highly specific and sensitive detection of 24 mRNA targets.
  • Demonstrated no significant compromise in sample variation across targets.
  • Confirmed the absence of inter-target biases in the amplification and detection process.
  • RT-isoPCR showed robust performance in multiplex mRNA analysis.

Conclusions:

  • RT-isoPCR is an effective method for sensitive and specific multiplex mRNA detection.
  • The technique minimizes biases, ensuring reliable quantification across multiple targets.
  • This approach offers a valuable tool for gene expression studies and diagnostic applications.