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

Next-generation Sequencing03:00

Next-generation Sequencing

100.8K
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....
100.8K
RNA-seq03:21

RNA-seq

12.4K
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...
12.4K
Real Time RT-PCR02:57

Real Time RT-PCR

66.4K
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...
66.4K
Sanger Sequencing01:57

Sanger Sequencing

777.9K
DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
777.9K

You might also read

Related Articles

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

Sort by
Same author

Wound Healing and Angiogenic Profiling of Dermal Endothelial Cells Isolated From People With Type 2 Diabetes.

FASEB journal : official publication of the Federation of American Societies for Experimental Biology·2026
Same author

ARNT2 repression disrupts neuronal identity and promotes glioblastoma growth.

Cell communication and signaling : CCS·2026
Same author

Decoding exercise adaptation through multidimensional biocircuitry.

Physiological genomics·2025
Same author

Role of tankyrase scaffolding in the β-catenin destruction complex and WNT signaling.

bioRxiv : the preprint server for biology·2025
Same author

Telomerase mRNA therapy protects human skin against radiation-induced DNA damage.

Molecular therapy : the journal of the American Society of Gene Therapy·2025
Same author

Wound healing and angiogenic profiling of dermal endothelial cells isolated from people with type 2 diabetes.

bioRxiv : the preprint server for biology·2025
Same journal

MT-MRI for detection of renal interstitial fibrosis in renovascular disease.

Scientific reports·2026
Same journal

Detection of underground objects from GPR data using a lightweight YOLO-based approach.

Scientific reports·2026
Same journal

Early systemic inflammatory-metabolic trajectory phenotypes are associated with survival outcomes in metastatic renal cell carcinoma treated with nivolumab.

Scientific reports·2026
Same journal

Water balance components in a dry-seeded rice-wheat system: Untangling the effects of tillage and mulching practices.

Scientific reports·2026
Same journal

Topological approaches to quantum tensor train compression via ZX-calculus and SVD.

Scientific reports·2026
Same journal

determinants of flood impacts and adaptive capacity among market vendors in Walukuba-Masese, Jinja city, Uganda.

Scientific reports·2026
See all related articles

Related Experiment Video

Updated: Mar 23, 2026

Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing
12:33

Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing

Published on: July 28, 2017

13.6K

Comparison of DNA Quantification Methods for Next Generation Sequencing.

Jérôme D Robin1, Andrew T Ludlow1, Ryan LaRanger1

  • 1Department of Cell Biology, 5323 Harry Hines Boulevard, UT Southwestern Medical Center, Dallas, TX 75390-9030, USA.

Scientific Reports
|April 7, 2016
PubMed
Summary
This summary is machine-generated.

Digital PCR (ddPCR) offers a precise method for quantifying Next Generation Sequencing (NGS) libraries, improving data quality. This study recommends ddPCR for accurate NGS library titration, overcoming limitations of traditional PCR-based methods.

More Related Videos

Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies
13:24

Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies

Published on: April 11, 2016

12.4K
Amplification of Near Full-length HIV-1 Proviruses for Next-Generation Sequencing
10:18

Amplification of Near Full-length HIV-1 Proviruses for Next-Generation Sequencing

Published on: October 16, 2018

12.8K

Related Experiment Videos

Last Updated: Mar 23, 2026

Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing
12:33

Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing

Published on: July 28, 2017

13.6K
Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies
13:24

Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies

Published on: April 11, 2016

12.4K
Amplification of Near Full-length HIV-1 Proviruses for Next-Generation Sequencing
10:18

Amplification of Near Full-length HIV-1 Proviruses for Next-Generation Sequencing

Published on: October 16, 2018

12.8K

Area of Science:

  • Genomics
  • Molecular Biology
  • Biotechnology

Background:

  • Next Generation Sequencing (NGS) is crucial for genomic studies, including cancer mutation analysis and chromatin interaction mapping.
  • Accurate quantification of NGS libraries is essential for reliable results, but a gold standard method is lacking.
  • Standard quantification methods often rely on PCR, which can introduce bias and affect rare variant detection.

Purpose of the Study:

  • To compare droplet digital PCR (ddPCR) technologies, including ddPCR-Tail, with standard methods for NGS library quantification.
  • To evaluate the accuracy and sensitivity of ddPCR for titrating NGS libraries.
  • To provide a comprehensive comparison of quantification techniques within a complete sequencing workflow.

Main Methods:

  • Comparison of droplet digital PCR (ddPCR) and ddPCR-Tail against traditional methods like qPCR and fluorometry (QuBit).
  • Assessment of quantification accuracy through analysis of barcode repartition after sequencing multiplexed samples.
  • Evaluation of library heterogeneity and rare variant detection capabilities.

Main Results:

  • ddPCR-Tail demonstrates comparable performance to qPCR and QuBit for NGS library quantification.
  • ddPCR allows sensitive quantification, particularly through barcode repartition analysis in multiplexed samples.
  • The study highlights potential improvements in NGS quality using ddPCR-based quantification.

Conclusions:

  • ddPCR-based quantification is a viable and sensitive alternative for NGS library titration.
  • Adopting ddPCR can enhance the accuracy and reliability of NGS experiments.
  • This research advocates for the integration of ddPCR for superior NGS quality control.