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 Videos

Allelic discrimination by denaturing high-performance liquid chromatography.

P J Oefner1

  • 1DNA Sequencing and Technology Center, Stanford University, Palo Alto, CA 94304, USA. oefner@genome.stanford.edu

Journal of Chromatography. B, Biomedical Sciences and Applications
|April 8, 2000
PubMed
Summary

This study introduces a high-performance liquid chromatography method for DNA analysis. It accurately distinguishes single-nucleotide polymorphisms (SNPs) in DNA, enabling rapid genetic variant genotyping.

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

BITES: balanced individual treatment effect for survival data.

Bioinformatics (Oxford, England)·2022
Same author

Molecular signatures that can be transferred across different omics platforms.

Bioinformatics (Oxford, England)·2017
Same author

Molecular signatures that can be transferred across different omics platforms.

Bioinformatics (Oxford, England)·2017
Same author

Reference point insensitive molecular data analysis.

Bioinformatics (Oxford, England)·2016
Same author

Rifaximin preserves intestinal microbiota balance in patients undergoing allogeneic stem cell transplantation.

Bone marrow transplantation·2016
Same author

Increased incidence of chronic GvHD and CMV disease in patients with vitamin D deficiency before allogeneic stem cell transplantation.

Bone marrow transplantation·2015

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Genetics

Background:

  • Single-nucleotide polymorphisms (SNPs) are key genetic variations.
  • Accurate and efficient genotyping methods are crucial for genetic research and diagnostics.
  • Current methods for SNP detection can be complex or require reference DNA.

Purpose of the Study:

  • To develop a novel high-performance liquid chromatography (HPLC) method for high-resolution DNA analysis.
  • To enable accurate allelic discrimination and genotyping of single-nucleotide polymorphisms (SNPs).
  • To provide a rapid genotyping method that does not require a reference chromosome.

Main Methods:

  • Utilizing ion-pair reversed-phase HPLC on specialized poly(styrene-divinylbenzene) particles.
  • Employing a preheated mobile phase for instantaneous complete denaturation of short DNA amplicons.

Related Experiment Videos

  • Analyzing DNA molecules differing by a single base, including transitions and transversions (excluding C-->G).

    • Main Results:

    • Achieved resolution of single-stranded DNA molecules of identical size (<100 nucleotides) that differ by a single base.
    • Demonstrated accurate typing of most single-nucleotide polymorphisms (SNPs) through allelic discrimination.
    • Successfully performed rapid genotyping of genetic variants without a reference chromosome.

    Conclusions:

    • Ion-pair reversed-phase HPLC offers a powerful tool for high-resolution DNA analysis and SNP genotyping.
    • The developed method complements existing SNP discovery techniques like heteroduplex analysis.
    • This approach facilitates rapid and accurate genetic variant genotyping, advancing applications in molecular diagnostics and research.