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 Experiment Video

Updated: May 20, 2026

DNA-based Fish Species Identification Protocol
09:15

DNA-based Fish Species Identification Protocol

Published on: April 28, 2010

A fast, highly sensitive double-nested PCR-based method to screen fish immunobiomes.

Sébastien Boutin1, Maelle Sevellec, Scott A Pavey

  • 1Institut de Biologie Intégrative et des Systèmes (IBIS), Département de Biologie, Université Laval, Québec, QC, Canada.

Molecular Ecology Resources
|July 19, 2012
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Multidisciplinary Evaluation of a 10-Year Restoration Program for Two Endangered Atlantic Salmon (<i>Salmo salar</i>) Populations.

Evolutionary applications·2026
Same author

Genome-Wide Population Structure of Lake Whitefish (<i>Coregonus clupeaformis</i>) in a Subarctic Great Lake.

Evolutionary applications·2026
Same author

Environmental RNA reflects seasonal changes in the metabolic activity of stream salamanders.

Scientific reports·2026
Same author

High-throughput multispecies quantitative polymerase chain reaction assays to study the effects of acute thermal stress in three species of Acipenser sturgeon.

Journal of fish biology·2026
Same author

Host genotype and environmental factors differentially shape the black piranha's gill microbiota.

Microbiology spectrum·2026
Same author

New Insights Into the Biogeography of Six <i>Garra</i> Species (Teleostei: Cyprinidae) in the Persian Gulf Basin.

Ecology and evolution·2026
Same journal

From Gene Copies to Cell Numbers: Advancing Quantitative Approaches in Protistan Ecology Using Digital PCR.

Molecular ecology resources·2026
Same journal

EasyCen: A Lightweight Framework for Centromere Localisation and Repeat-Organisation Profiling in Telomere-to-Telomere Genomes.

Molecular ecology resources·2026
Same journal

A Practical Framework for GT-Seq Panel Optimization.

Molecular ecology resources·2026
Same journal

Comparison of Environmental DNA and Bulk DNA Metabarcoding for Assessing Terrestrial Arthropod Diversity Across Three Habitat Types on Guam.

Molecular ecology resources·2026
Same journal

pr2-Wormifier: A Bioinformatics Pipeline to Create Custom Reference Databases for Improved Metabarcoding of Marine Protists.

Molecular ecology resources·2026
Same journal

Individual Identification of Prey in Carnivore Scats.

Molecular ecology resources·2026
See all related articles

A new double-nested PCR method efficiently detects infectious bacteria in host tissues. This technique is cost-effective for screening low-density microbiomes, like fish immunobiomes, using minimal samples.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Aquatic Animal Health

Background:

  • Screening bacterial diversity in host tissues requires efficient molecular methods.
  • Existing PCR techniques struggle with low bacterial loads and specificity.
  • Accurate assessment of infectious agents is crucial for host health.

Purpose of the Study:

  • To develop and validate a double-nested PCR methodology for constructing 16S tag amplicon libraries.
  • To assess the efficiency and specificity of this method for detecting bacterial DNA from host samples.
  • To evaluate its applicability for screening infectious bacterial diversity in fish immunobiomes.

Main Methods:

  • Developed a double-nested PCR protocol for 16S rRNA gene amplification.
  • Applied the method to 133 lake whitefish kidney samples.

More Related Videos

Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis
06:30

Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis

Published on: February 5, 2014

Immunization of Adult Zebrafish for the Preclinical Screening of DNA-based Vaccines
05:39

Immunization of Adult Zebrafish for the Preclinical Screening of DNA-based Vaccines

Published on: October 30, 2018

Related Experiment Videos

Last Updated: May 20, 2026

DNA-based Fish Species Identification Protocol
09:15

DNA-based Fish Species Identification Protocol

Published on: April 28, 2010

Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis
06:30

Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis

Published on: February 5, 2014

Immunization of Adult Zebrafish for the Preclinical Screening of DNA-based Vaccines
05:39

Immunization of Adult Zebrafish for the Preclinical Screening of DNA-based Vaccines

Published on: October 30, 2018

  • Compared double-nested PCR with single-pair and simple nested PCR strategies.
  • Utilized 454 FLX pyrosequencing for amplicon library analysis.

    • Main Results:

    • Double-nested PCR demonstrated highly specific amplification of bacterial DNA, outperforming other methods.
    • Achieved high sequencing coverage (mean Good's coverage = 95.4%) with minimal chimeric amplicons.
    • Identified variations in pathogenic bacteria abundance within fish populations.

    Conclusions:

    • The double-nested PCR approach is a rapid, informative, and cost-effective method for screening fish immunobiomes.
    • This methodology is highly effective for analyzing bacterial diversity even with very small sample amounts.
    • The technique shows promise for application to other low-density microbiome studies.