Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

RNA-seq03:21

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
Ribosome Profiling02:24

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Next-generation Sequencing03:00

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.

You might also read

Related Articles

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

Sort by
Same author

Resolving the Haplotype Complexity of Colorectal Cancer Genomes with Droplet Barcode Sequencing.

Life (Basel, Switzerland)·2026
Same author

Diagnostic Factors Associated with Sarcoidosis in Patients Referred for EBUS-TBNA Due to Mediastinal Lymphadenopathy.

Advances in respiratory medicine·2026
Same author

Survival and Treatment Modalities in Primary Vaginal Melanoma-Case Report and a Narrative Review.

Journal of clinical medicine·2024
Same author

BLR: a flexible pipeline for haplotype analysis of multiple linked-read technologies.

Nucleic acids research·2023
Same author

Characterizing single extracellular vesicles by droplet barcode sequencing for protein analysis.

Journal of extracellular vesicles·2022
Same author

High throughput barcoding method for genome-scale phasing.

Scientific reports·2019
Same journal

Turbulent flow in a vortex separator with a directed pipe inlet.

Scientific reports·2026
Same journal

Systematic characteristic evaluation of clay-based cementitious material derived from calcium carbide residue and waste tile powder.

Scientific reports·2026
Same journal

Retraction Note: Improvement of a rapid diagnostic application of monoclonal antibodies against avian influenza H7 subtype virus using Europium nanoparticles.

Scientific reports·2026
Same journal

Applying large language models to spam detection in the Kazakh low-resource language setting.

Scientific reports·2026
Same journal

An open-source 3D printing system enabling in-situ freeze-thaw processing of hydrogels.

Scientific reports·2026
Same journal

An enhanced EfficientNet framework for automated waste classification using cosine annealing and label smoothing.

Scientific reports·2026
See all related articles

Related Experiment Video

Updated: May 17, 2026

Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations
11:52

Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations

Published on: August 4, 2016

Targeted transcript profiling by sequencing.

Pawel Zajac1, Afshin Ahmadian

  • 1Laboratory for Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.

Scientific Reports
|November 10, 2012
PubMed
Summary
This summary is machine-generated.

This study introduces trinucleotide threading (TnT) amplification with massive parallel sequencing for accurate gene expression profiling. This method reliably measures gene expression even with low RNA input, minimizing the impact of unwanted products.

More Related Videos

Comprehensive Spatial Profiling of Species-agnostic Transcriptomes via Stereo-seq
10:22

Comprehensive Spatial Profiling of Species-agnostic Transcriptomes via Stereo-seq

Published on: October 31, 2025

Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling
10:00

Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling

Published on: October 28, 2014

Related Experiment Videos

Last Updated: May 17, 2026

Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations
11:52

Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations

Published on: August 4, 2016

Comprehensive Spatial Profiling of Species-agnostic Transcriptomes via Stereo-seq
10:22

Comprehensive Spatial Profiling of Species-agnostic Transcriptomes via Stereo-seq

Published on: October 31, 2025

Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling
10:00

Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling

Published on: October 28, 2014

Area of Science:

  • Molecular Biology
  • Genomics
  • Gene Expression Analysis

Background:

  • Trinucleotide threading (TnT) amplification combined with array readout accurately monitors gene expression levels.
  • Array-based detection of TnT products can fail to identify spurious amplification products, potentially affecting data accuracy.
  • Massive parallel sequencing offers an unbiased approach to analyze the complete TnT-generated product population.

Purpose of the Study:

  • To adapt the TnT protocol for massive parallel sequencing to obtain an unbiased view of all generated products.
  • To investigate the identity and prevalence of undesired products in TnT amplification.
  • To assess the impact of undesired products on gene expression level measurements at varying RNA input concentrations.

Main Methods:

  • Targeted gene expression profiling using trinucleotide threading (TnT) amplification.
  • Massive parallel sequencing for unbiased readout of TnT-generated products.
  • Analysis of undesired product formation across different oligonucleotide:RNA ratios.

Main Results:

  • TnT gene expression profiling with massive sequencing provides reliable data from as little as 3.5 ng of total RNA.
  • Using 350 ng of total RNA resulted in only 0.7% to 1.1% undesired products.
  • Decreasing input RNA increased the fraction of undesired products, but expression profiles remained unaffected.

Conclusions:

  • Massive parallel sequencing is a robust readout strategy for TnT amplification, enabling accurate gene expression profiling.
  • The TnT method with sequencing is reliable even with limited total RNA input.
  • Undesired products generated during TnT amplification do not compromise the accuracy of expression profiles, even at lower RNA input levels.