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PCR

PCR

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: Jul 6, 2026

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Published on: March 30, 2015

MultiPriDe: automated batch development of quantitative real-time PCR primers.

A C Ziesel¹, M A Chrenek, P W Wong

¹Department of Ophthalmology, Emory University, B5500 Clinic B, 1365 Clifton Road NE, Atlanta, GA 30322, USA. aziesel@emory.edu

Nucleic Acids Research

|April 11, 2008

Summary

Designing primers for quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) is now more efficient. MultiPriDe automates primer design for multiple genes, reducing errors and improving gene expression analysis.

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Last Updated: Jul 6, 2026

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Published on: March 30, 2015

Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR (qPCR) Arrays

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Published on: December 3, 2010

Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp

Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp

Published on: June 20, 2018

Area of Science:

Molecular Biology
Bioinformatics

Background:

Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) is a standard method for gene expression analysis.
Developing specific oligonucleotide primers is crucial but often manual and error-prone for large-scale studies.

Purpose of the Study:

To develop an automated tool for efficient and accurate primer design for qRT-PCR.
To streamline the process of identifying optimal primer pairs for multiple genes.

Main Methods:

Developed MultiPriDe (Multiple Primer Design), a Perl utility for batch primer design.
Integrated MultiPriDe with the Primer3 utility, utilizing gene-specific intron/exon data.
Incorporated optimized hybridization conditions to enhance primer design success.

Main Results:

MultiPriDe automates the identification of target sites and primer generation for multiple genes.
The utility enhances efficiency and reduces user error in the primer design workflow.
Facilitates the management of primer information for large gene sets.

Conclusions:

MultiPriDe significantly improves the efficiency of primer design for qRT-PCR.
The automated approach aids researchers in large-scale gene expression studies.
Offers a robust solution for managing primer design challenges.