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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: Apr 29, 2026

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

Methods for multiplex template sampling in digital PCR assays.

Oleh I Petriv¹, Kevin A Heyries¹, Michael VanInsberghe¹

¹Centre for High-Throughput Biology, University of British Columbia, Vancouver, British Columbia, Canada.

|May 24, 2014

Summary

This summary is machine-generated.

Multiplex Template Sampling (MTS) enhances digital PCR (dPCR) precision by increasing target molecule concentrations. This method effectively boosts template levels from fragmented molecules, crucial for accurate copy number analysis in clinical samples.

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Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

Digital PCR (dPCR) offers precise copy number analysis.
High target molecule concentrations are essential for dPCR efficiency.
Clinical samples often yield insufficient target molecule concentrations.

Purpose of the Study:

To develop a method for increasing template concentrations for dPCR.
To enable precise copy number analysis from limited clinical samples.
To present Multiplex Template Sampling (MTS) as a solution.

Main Methods:

Multiplex Template Sampling (MTS) strategy.
Detecting multiple regions of fragmented target molecules.
Developing three alternative assay approaches for MTS analysis of chromosome 21.

Main Results:

Achieved a 10-fold concentration enhancement.
Preserved assay precision despite increased concentration.
Demonstrated feasibility of MTS for chromosome 21 analysis.

Conclusions:

MTS effectively increases template concentrations for dPCR.
MTS is a viable strategy for improving precision in copy number analysis.
This method addresses limitations of low-input clinical samples for dPCR.