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

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.
Sanger Sequencing01:57

Sanger Sequencing

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...
The Replisome03:01

The Replisome

DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
Lagging Strand Synthesis01:59

Lagging Strand Synthesis

During replication, the complementary strands in double-stranded DNA are synthesized at different rates. Replication first begins on the leading strand. Replication starts later, occurs more slowly, and proceeds discontinuously on the lagging strand.
There are several major differences between synthesis of the leading strand and synthesis of the lagging strand. 1) Leading strand synthesis happens in the direction of replication fork opening, whereas lagging strand synthesis happens in the...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...

You might also read

Related Articles

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

Sort by
Same author

CRISPR/Cas9-mediated genome editing reveals the involvement of a polyphenol oxidase in the shikonin-specific biosynthesis in Lithospermum erythrorhizon.

Plant & cell physiology·2026
Same author

Peptidomics and Beyond: Development of a Novel Platform Based on Capillary Electrophoresis-Mass Spectrometry for Metabolomics of Medium-Molecular-Weight Compounds.

Journal of proteome research·2026
Same author

Revisiting the Oxidation State of [Cu(HIO<sub>6</sub>)<sub>2</sub>]<sup>5-</sup>: A Copper(III) Complex between Classical Werner-Type and Inverted Ligand Fields.

Inorganic chemistry·2026
Same author

S-alk(en)yl-cysteine sulfoxides in Allium species are excellent acrolein scavengers: implications for secondary antioxidants in plants.

Bioscience, biotechnology, and biochemistry·2025
Same author

Activation of Anionic Redox for Stoichiometric and Li-Excess Metal Sulfides through Structural Disordering: Joint Experimental and Theoretical Study.

Journal of the American Chemical Society·2025
Same author

Photochromic Color Tuning of Copper-Doped Zinc Sulfide Nanocrystals by Control of Local Dopant Environments.

Angewandte Chemie (International ed. in English)·2025
Same journal

Increased abundance of Limosilactobacillus reuteri in the gut of selectively bred high-tameness mice and its association with behavioural changes.

DNA research : an international journal for rapid publication of reports on genes and genomes·2026
Same journal

PFGPred: a stack ensemble classifier for the identification of fusion genes in plants.

DNA research : an international journal for rapid publication of reports on genes and genomes·2026
Same journal

AlleleMiner: a long-read pipeline for gene-wise de novo allele phasing and variant detection in diploid citrus cultivars.

DNA research : an international journal for rapid publication of reports on genes and genomes·2026
Same journal

Silver chimaera genome highlights lineage-specific sex chromosome and opsin gene evolution in holocephalans.

DNA research : an international journal for rapid publication of reports on genes and genomes·2026
Same journal

Chromosome-level genome assembly of the Gerbera (Gerbera hybrida) using HiFi long-read and Hi-C technologies.

DNA research : an international journal for rapid publication of reports on genes and genomes·2026
Same journal

Chromosome-scale genome assembly of Malcolmia littorea using long-read sequencing and single-pollen genotyping technologies.

DNA research : an international journal for rapid publication of reports on genes and genomes·2026
See all related articles

Related Experiment Video

Updated: May 9, 2026

Using Modified Synthetic Oligonucleotides to Assay Nucleic Acid-Metabolizing Enzymes
05:33

Using Modified Synthetic Oligonucleotides to Assay Nucleic Acid-Metabolizing Enzymes

Published on: July 5, 2024

Precise sequential DNA ligation on a solid substrate: solid-based rapid sequential ligation of multiple DNA

Eiji Takita1, Katsunori Kohda, Hajime Tomatsu

  • 11 Kazusa DNA Research Institute, 2-6-7 Kazusa-kamatari, Kisarazu, Chiba 292-0818, Japan.

DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes
|July 31, 2013
PubMed
Summary
This summary is machine-generated.

Researchers developed PRESSO, a novel method for precise sequential DNA ligation on a solid substrate. This technique efficiently joins multiple DNA fragments in tandem, advancing genetic engineering and functional genomics.

Keywords:
DNA ligationcloningfunctional genomicsmultiple DNA fragment assembly

More Related Videos

Automated Robotic Liquid Handling Assembly of Modular DNA Devices
11:22

Automated Robotic Liquid Handling Assembly of Modular DNA Devices

Published on: December 1, 2017

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage
06:51

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

Published on: May 6, 2020

Related Experiment Videos

Last Updated: May 9, 2026

Using Modified Synthetic Oligonucleotides to Assay Nucleic Acid-Metabolizing Enzymes
05:33

Using Modified Synthetic Oligonucleotides to Assay Nucleic Acid-Metabolizing Enzymes

Published on: July 5, 2024

Automated Robotic Liquid Handling Assembly of Modular DNA Devices
11:22

Automated Robotic Liquid Handling Assembly of Modular DNA Devices

Published on: December 1, 2017

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage
06:51

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

Published on: May 6, 2020

Area of Science:

  • Molecular Biology
  • Genetic Engineering
  • Functional Genomics

Background:

  • DNA ligation is crucial for molecular cloning and producing genetically modified organisms.
  • Tandemly incorporating multiple gene cassettes is often necessary for generating organisms with multiple foreign gene products.

Purpose of the Study:

  • To introduce a novel method, PRESSO (precise sequential DNA ligation on a solid substrate), for efficient tandem ligation of multiple DNA fragments.
  • To overcome technical challenges in functional genomics related to generating transgenic constructs with multiple genes.

Main Methods:

  • Donor DNA fragments were amplified with non-palindromic ends.
  • Fragments were ligated to acceptor DNA fragments immobilized on solid beads.
  • The final construct was released from beads using rare-cut meganuclease and circularized via intramolecular ligation.

Main Results:

  • The PRESSO method enabled rapid and efficient joining of multiple genes in a desired order and orientation.
  • Demonstrated successful tandem ligation of DNA fragments on a solid substrate.

Conclusions:

  • PRESSO offers a robust solution for constructing complex DNA molecules with multiple genes.
  • This method significantly advances the generation of transgenic organisms for research and applied biosciences.