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PCR

PCR

PCR - Polymerase Chain Reaction

PCR - Polymerase Chain Reaction

Real Time RT-PCR

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: Jun 7, 2026

Detection of Human Immunodeficiency Virus Type 1 (HIV-1) Antisense Protein (ASP) RNA Transcripts in Patients by Strand-Specific RT-PCR

Detection of Human Immunodeficiency Virus Type 1 (HIV-1) Antisense Protein (ASP) RNA Transcripts in Patients by Strand-Specific RT-PCR

Published on: November 27, 2019

Asynchronous PCR.

Caifu Chen¹, David Ruff, Jason Halsey

¹Genomic Assays R&D, Molecular Biology Systems Division, Life Technologies, Foster City, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)

|October 23, 2010

Summary

This summary is machine-generated.

Asynchronous PCR (aPCR) enables ordered DNA amplification using distinct primer melting temperatures and concentrations. This method enhances hybridization probe binding for applications like gene expression analysis.

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Area of Science:

Molecular Biology
Biotechnology

Background:

Traditional PCR amplifies both DNA strands simultaneously.
Existing methods may not optimally generate single-stranded DNA for hybridization assays.

Purpose of the Study:

Introduce and characterize Asynchronous PCR (aPCR) for ordered DNA amplification.
Explore aPCR's potential in enhancing hybridization-based applications.

Main Methods:

Utilizing forward and reverse primers with a melting temperature difference of at least 15°C.
Reducing the concentration of the lower melting temperature primer to 100 nM.
Implementing a unique thermocycling protocol with two annealing and extension steps per cycle.

Main Results:

Achieved ordered and sequential amplification of DNA strands.
Generated transient single-stranded DNA (ssDNA) amplicons.
Demonstrated improved binding of hybridization probes, such as peptide nucleic acids (PNAs).

Conclusions:

aPCR offers a novel approach for strand-specific DNA amplification.
The method facilitates robust ssDNA production for hybridization applications.
aPCR is suitable for real-time quantitative PCR (qPCR) and microarray analyses.