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

10.6K
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...
10.6K
Next-generation Sequencing03:00

Next-generation Sequencing

94.2K
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....
94.2K

You might also read

Related Articles

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

Sort by
Same author

Elevated cfDNA after exercise is derived primarily from mature polymorphonuclear neutrophils, with a minor contribution of cardiomyocytes.

Cell reports. Medicine·2026
Same author

Disruption of the brain-spleen axis impairs monocyte-microglia communication and accelerates disease progression in a mouse model of amyloidosis.

Nature communications·2026
Same author

Cell-to-cell variability and gain of methylation at polycomb CpG islands as a hallmark of aging.

Nature communications·2026
Same author

Beyond Circulating Tumor DNA for Efficacy: Can We Use Cell-Free DNA to Detect and Monitor Toxicity Signals?

JCO precision oncology·2026
Same author

The genetic basis for DNA methylation variation across tissues and development.

Nature communications·2026
Same author

Plasma cell-free DNA markers predict occult metastases in patients with resectable pancreatic ductal adenocarcinoma.

Clinical and translational medicine·2026

Related Experiment Video

Updated: Oct 18, 2025

Large-Scale SARS-CoV-2 Testing Utilizing Saliva and Transposition Sample Pooling
08:26

Large-Scale SARS-CoV-2 Testing Utilizing Saliva and Transposition Sample Pooling

Published on: June 23, 2022

1.9K

Early sample tagging and pooling enables simultaneous SARS-CoV-2 detection and variant sequencing.

Alon Chappleboim1,2, Daphna Joseph-Strauss1,2, Ayelet Rahat1,2

  • 1Alexander Silberman Institute of Life Science, Hebrew University of Jerusalem, Jerusalem 9190401, Israel.

Science Translational Medicine
|September 30, 2021
PubMed
Summary

ApharSeq offers a streamlined approach for diagnosing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using next-generation sequencing (NGS). This method significantly reduces costs and labor for high-throughput testing, providing accurate variant information.

More Related Videos

Detection of Rare Genomic Variants from Pooled Sequencing Using SPLINTER
14:06

Detection of Rare Genomic Variants from Pooled Sequencing Using SPLINTER

Published on: June 23, 2012

15.4K
Quantification and Whole Genome Characterization of SARS-CoV-2 RNA in Wastewater and Air Samples
09:26

Quantification and Whole Genome Characterization of SARS-CoV-2 RNA in Wastewater and Air Samples

Published on: June 30, 2023

1.3K

Related Experiment Videos

Last Updated: Oct 18, 2025

Large-Scale SARS-CoV-2 Testing Utilizing Saliva and Transposition Sample Pooling
08:26

Large-Scale SARS-CoV-2 Testing Utilizing Saliva and Transposition Sample Pooling

Published on: June 23, 2022

1.9K
Detection of Rare Genomic Variants from Pooled Sequencing Using SPLINTER
14:06

Detection of Rare Genomic Variants from Pooled Sequencing Using SPLINTER

Published on: June 23, 2012

15.4K
Quantification and Whole Genome Characterization of SARS-CoV-2 RNA in Wastewater and Air Samples
09:26

Quantification and Whole Genome Characterization of SARS-CoV-2 RNA in Wastewater and Air Samples

Published on: June 30, 2023

1.3K

Area of Science:

  • Molecular diagnostics
  • Virology
  • Genomic sequencing

Background:

  • Current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnostics primarily use RNA extraction and RT-qPCR.
  • Next-generation sequencing (NGS) offers higher throughput and variant information but is currently costly and complex.
  • Existing NGS assays for SARS-CoV-2 involve multiple parallel sample processing steps, increasing expenses.

Purpose of the Study:

  • To develop and validate ApharSeq, a cost-effective and efficient NGS-based diagnostic assay for SARS-CoV-2.
  • To enable simultaneous detection and genotyping of SARS-CoV-2 with high throughput.
  • To reduce labor and reagent costs in large-scale diagnostic testing.

Main Methods:

  • ApharSeq involves barcoding samples in lysis buffer, followed by pooling before reverse transcription.
  • The assay was validated on over 500 clinical samples using a robotic workflow.
  • Unique molecular identifiers were incorporated for high-confidence genotype determination.

Main Results:

  • ApharSeq demonstrated linearity across five orders of magnitude with a limit of detection at Ct 33 (95% sensitivity) and >99.5% specificity.
  • The method provided targeted, high-confidence genotype information.
  • Early pooling in ApharSeq led to an estimated 10- to 100-fold reduction in labor, automation, and reagent use for high-throughput settings.

Conclusions:

  • ApharSeq presents a promising, cost-effective NGS diagnostic tool for SARS-CoV-2.
  • The protocol's flexibility allows for simultaneous assaying of other RNA targets.
  • ApharSeq is well-suited for addressing current and future mass diagnostic challenges efficiently.