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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: Oct 10, 2025

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

Published on: March 31, 2021

An Open One-Step RT-qPCR for SARS-CoV-2 detection.

Ariel Cerda^1,2, Maira Rivera^1,3, Grace Armijo^1,2

¹ANID - Millennium Science Initiative Program - Millennium Institute for Integrative Biology (iBio).

Medrxiv : the Preprint Server for Health Sciences

|December 15, 2021

Summary

A new open-source protocol for detecting SARS-CoV-2 RNA using One-Step RT-qPCR offers a cost-effective alternative. This approach ensures accessibility for diagnostic testing in resource-limited settings globally.

Keywords:

Diagnostic system One-Step RT-PCR Open Source Recombinant enzymes SARS-CoV-2

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Last Updated: Oct 10, 2025

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

Published on: March 31, 2021

Author Spotlight: Advancements in Multiplex Detection of Respiratory Viruses

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Author Spotlight: Advancing Pathogen Diagnostics with Standardized LAMP

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Published on: September 8, 2023

Area of Science:

Molecular Biology
Virology
Biotechnology

Background:

The COVID-19 pandemic highlighted global reliance on RT-PCR diagnostics.
Centralized reagent production and IP restrictions hinder access in developing nations.
Development of accessible, standardized diagnostic protocols is crucial.

Approach:

Established a One-Step open RT-qPCR protocol for SARS-CoV-2 RNA detection.
Purified and validated recombinant M-MLV RT and Taq DNA pol enzymes.
Utilized a non-proprietary buffer and Taqman probe-based assay.

Key Points:

The open RT-qPCR protocol demonstrated comparable sensitivity and performance to commercial kits.
Validation was performed using synthetic RNA and clinical samples.
The protocol utilizes readily available, non-proprietary components.

Conclusions:

This study presents a proof of concept for an open-access diagnostic approach.
The developed protocol offers a potentially cost-effective and accessible solution for viral diagnostics.
This open approach can improve global health equity in infectious disease detection.