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

A rapid method for purifying PCR products for direct sequence analysis.

M H Kalnoski1, M F McCoy-Haman, G F Hollis

  • 1Department of Biological Sciences, Monsanto Company, St. Louis, MO 63198.

Biotechniques
|August 1, 1991
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

Plasmid construction by linker-assisted homologous recombination in yeast.

BioTechniques·2002
Same author

A comprehensive system for protein purification and biochemical analysis based on antibodies to c-myc peptide.

Protein expression and purification·2001
Same author

Unusually stable and long-lived ligand-induced conformations of integrins.

The Journal of biological chemistry·2001
Same author

Aspartyl beta -hydroxylase (Asph) and an evolutionarily conserved isoform of Asph missing the catalytic domain share exons with junctin.

The Journal of biological chemistry·2000
Same author

A yeast genetic assay for caspase cleavage of the amyloid-beta precursor protein.

Molecular biotechnology·2000
Same author

A counterselection for the tryptophan pathway in yeast: 5-fluoroanthranilic acid resistance.

Yeast (Chichester, England)·2000

This study presents a fast HPLC method for purifying and sequencing double-stranded DNA directly from PCR. The technique offers a simple, reliable way to obtain DNA sequences from small samples efficiently.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Polymerase Chain Reaction (PCR) generates DNA fragments for analysis.
  • Purification of PCR products is often a prerequisite for sequencing.
  • Existing purification methods can be time-consuming or involve organic solvents.

Purpose of the Study:

  • To describe a novel High-Performance Liquid Chromatography (HPLC) method for DNA purification and sequencing.
  • To offer a rapid, efficient, and reliable alternative to conventional DNA purification techniques.
  • To demonstrate the method's utility in sequencing specific gene fragments.

Main Methods:

  • Development of an HPLC-based protocol for direct purification of double-stranded DNA from PCR reactions.
  • Elution of purified DNA fragments within a 7-minute timeframe.

Related Experiment Videos

  • Sequencing of purified DNA fragments to assess the method's accuracy and yield.
  • Application of the method to sequence the tumor necrosis factor alpha (TNF alpha) gene.

    • Main Results:

    • Successful purification and sequencing of double-stranded DNA directly from PCR.
    • Rapid DNA elution time of under 7 minutes.
    • No requirement for organic cleanup steps.
    • Generation of several hundred bases of sequence data.
    • High sensitivity, enabling DNA sequencing from a single 100-microliter PCR reaction.

    Conclusions:

    • The described HPLC approach is a simple, reliable, and rapid method for DNA purification and sequencing.
    • This technique eliminates the need for organic solvents, simplifying the workflow.
    • The method is sensitive and efficient for obtaining DNA sequence information from PCR amplicons.