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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: Jun 24, 2025

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Published on: June 8, 2019

Ultra-Rapid Droplet Digital PCR Enables Intraoperative Tumor Quantification.

Zachary R Murphy^1,2, Emilia C Bianchini^1,2, Andrew Smith³

¹Center for Human Genetics and Genomics, New York University Grossman School of Medicine, USA.

Medrxiv : the Preprint Server for Health Sciences

|June 10, 2024

Summary

Ultra-Rapid droplet digital PCR (UR-ddPCR) provides molecular-genetic data in 15 minutes during surgery. This rapid molecular profiling aids intraoperative tumor diagnosis and guides surgical resections for improved patient outcomes.

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Last Updated: Jun 24, 2025

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

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Published on: June 8, 2019

Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR

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Published on: June 26, 2016

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Published on: September 25, 2018

Area of Science:

Oncology
Molecular Diagnostics
Surgical Pathology

Background:

Tumor diagnosis and treatment rely on molecular-genetic data.
Intraoperative molecular data access is limited, hindering real-time surgical decisions.
Current methods lack the speed for immediate intraoperative molecular analysis.

Purpose of the Study:

To develop a rapid molecular diagnostic tool for intraoperative use.
To enable real-time molecular profiling during surgery.
To improve tumor margin assessment and surgical guidance.

Main Methods:

Development of Ultra-Rapid droplet digital PCR (UR-ddPCR).
Demonstration of UR-ddPCR assays for IDH1 R132H and BRAF V600E mutations.
Intraoperative application in 13 glioma cases.
Integration with Ultra-Rapid stimulated Raman histology for cell density estimation.

Main Results:

UR-ddPCR delivers results in 15 minutes with accuracy comparable to standard ddPCR.
Successful intraoperative application in glioma patients.
Combined approach estimates both tumor cell percentages and densities.
Feasibility of point-of-care molecular diagnostics demonstrated.

Conclusions:

UR-ddPCR offers rapid, accurate molecular data during surgery.
This technology facilitates molecularly-guided surgeries.
Future refinements may enhance point-of-care diagnostics and clinical outcomes.