Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Restoring DSCAM expression mitigates neuronal morphology and axon guidance deficits in Down syndrome.

Brain : a journal of neurology·2026

Same author

Appendicectomy: a disease-modifying treatment for ulcerative colitis?

The lancet. Gastroenterology & hepatology·2026

Same author

Early-life dentine-based elemental biodynamics and cord blood telomere length.

medRxiv : the preprint server for health sciences·2026

Same author

Intestinal Inflammation Impacts Gestational Weight Gain in Women With Inflammatory Bowel Disease and Growth in Offspring.

Clinical gastroenterology and hepatology : the official clinical practice journal of the American Gastroenterological Association·2026

Same author

Multiple mTOR RNA localization signals regulate subcellular protein synthesis and axonal growth.

bioRxiv : the preprint server for biology·2026

Same author

Human induced pluripotent stem cell-based models for studying neural repair.

Progress in neurobiology·2026

Same journal

A Video Protocol of a Randomized Controlled Clinical Trial - Electrochemotherapy of Cutaneous Metastases with Reduced Dose Bleomycin (BLESS Trial).

Journal of visualized experiments : JoVE·2026

Same journal

A Standardized Ex Vivo Porcine Oromucosal Model for Evaluating Peptide Fluxes.

Journal of visualized experiments : JoVE·2026

Same journal

Lightweight English Text Classification with Deep Learning Based on Complex System Theory.

Journal of visualized experiments : JoVE·2026

Same journal

Integrating Artificial Intelligence-Assisted Translation Support into English Courses: Effects on Translation Accuracy, Perceived Stress, and Anxiety.

Journal of visualized experiments : JoVE·2026

Same journal

A Toxin-Based Counter-Selection System for Markerless Gene Deletion and High-Density Tn5 Transposon Mutagenesis in Pectobacterium brasiliense.

Journal of visualized experiments : JoVE·2026

Same journal

Seamless Multimodal Human-Robot Communication: Integration Techniques in Human-Computer Interaction.

Journal of visualized experiments : JoVE·2026

See all related articles

Search research articles

Related Experiment Video

Updated: May 5, 2026

RNA Isolation from Cell Specific Subpopulations Using Laser-capture Microdissection Combined with Rapid Immunolabeling

RNA Isolation from Cell Specific Subpopulations Using Laser-capture Microdissection Combined with Rapid Immunolabeling

Published on: April 11, 2015

Isolation and Quantification of Axonal mRNAs Using Porous Membrane Inserts and RTddPCR.

Shruti Ghumra¹, Manasi Agrawal¹, Meghal Desai¹

¹Department of Biological Sciences, Rutgers University- Newark.

Journal of Visualized Experiments : Jove

|February 23, 2026

Summary

This summary is machine-generated.

Researchers developed a new method to separate and quantify mRNA in neuronal compartments. This technique precisely measures localized RNA in axons, aiding the study of neuronal function and disease.

More Related Videos

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

Isolation and Quantification of Axonal mRNAs Using Porous Membrane Inserts and RTddPCR

Isolation and Quantification of Axonal mRNAs Using Porous Membrane Inserts and RTddPCR

Published on: February 6, 2026

Related Experiment Videos

Last Updated: May 5, 2026

RNA Isolation from Cell Specific Subpopulations Using Laser-capture Microdissection Combined with Rapid Immunolabeling

RNA Isolation from Cell Specific Subpopulations Using Laser-capture Microdissection Combined with Rapid Immunolabeling

Published on: April 11, 2015

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

Isolation and Quantification of Axonal mRNAs Using Porous Membrane Inserts and RTddPCR

Isolation and Quantification of Axonal mRNAs Using Porous Membrane Inserts and RTddPCR

Published on: February 6, 2026

Area of Science:

Neuroscience
Molecular Biology
Genetics

Background:

Neuronal function relies on localized mRNA and translation, particularly in axons.
Quantifying subcellular RNA populations in neurons is technically challenging.
Axonal mRNA localization is critical for neuronal connectivity, plasticity, and injury response.

Purpose of the Study:

To present a reproducible method for separating somatic and axonal compartments in cultured neurons.
To enable precise quantification of compartment-specific mRNA expression.
To facilitate the study of axonal RNA dynamics in response to various stimuli.

Main Methods:

Culturing rodent neurons on porous membranes (1-3 µm) to physically separate somatic and axonal compartments.
Isolating RNA from whole neuron and axon-enriched fractions.
Utilizing reverse transcriptase droplet digital PCR (RTddPCR) for high-sensitivity, absolute quantification of mRNA.

Main Results:

Achieved reproducible separation of somatic and axonal neuronal compartments.
Enabled absolute quantification of mRNA in distinct subcellular locations.
Demonstrated high sensitivity and reproducibility in detecting low-copy-number axonal transcripts.
Validated the method for studying RNA changes over time and in response to stimuli.

Conclusions:

The developed method provides a flexible, sensitive, and reproducible approach for studying axonal mRNA localization.
This technique allows for precise measurement of steady-state RNA abundance and changes in response to neurotrophic factors, stress, or injury.
The method supports downstream applications like protein synthesis measurement and perturbation experiments, advancing understanding of neuronal function and disease mechanisms.