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Related Concept Videos

Genome Annotation and Assembly03:36

Genome Annotation and Assembly

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The genome refers to all of the genetic material in an organism. It can range from a few million base pairs in microbial cells to several billion base pairs in many eukaryotic organisms. Genome assembly refers to the process of taking the DNA sequencing data and putting it all back together in a correct order to create a close representation of the original genome. This is followed by the identification of functional elements on the newly assembled genome, a process called genome annotation.
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DNA as a Genetic Template02:05

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Two structural features of the DNA molecule provide a basis for the mechanisms of heredity: the four nucleotide bases and its double-stranded nature. The Watson-Crick model of double-helical DNA structure, proposed in 1952, drew heavily upon the X-ray crystallography work of researchers Rosalind Franklin and Maurice Wilkins. Watson, Crick, and Wilkins jointly received the Nobel Prize in Physiology or Medicine for their work in 1962. Franklin was, controversially, excluded from the prize for...
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Sanger Sequencing01:57

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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...
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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.
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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.
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Genome comparison is one of the excellent ways to interpret the evolutionary relationships between organisms. The basic principle of genome comparison is that if two species share a common feature, it is likely encoded by the DNA sequence conserved between both species. The advent of genome sequencing technologies in the late 20th century enabled scientists to understand the concept of conservation of domains between species and helped them to deduce evolutionary relationships across diverse...
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Updated: Sep 18, 2025

Automated Robotic Liquid Handling Assembly of Modular DNA Devices
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UniClo: scarless hierarchical DNA assembly without sequence constraint.

Carol N Flores-Fernández1, Da Lin1, Katherine Robins1

  • 1Centre for Human Genetics, Nuffield Department of Medicine, University of Oxford, Oxford OX3 7BN, United Kingdom.

Nucleic Acids Research
|June 23, 2025
PubMed
Summary
This summary is machine-generated.

UniClo offers a universal and flexible DNA assembly method overcoming sequence constraints and scars. This novel approach uses DNA methylation and CRISPR-dCas9 technology for scarless, multi-round DNA fragment assembly.

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Area of Science:

  • Molecular Biology
  • Synthetic Biology
  • Biotechnology

Background:

  • Type IIs restriction enzyme-mediated DNA assembly is efficient but limited by sequence constraints and scar formation.
  • Existing methods often require complex designs and can introduce unwanted sequence artifacts.
  • There is a need for versatile and scarless DNA assembly techniques in molecular biology.

Purpose of the Study:

  • To develop a universal and flexible DNA assembly method that overcomes the limitations of traditional Type IIs restriction enzyme-mediated approaches.
  • To introduce a scarless DNA assembly strategy that simplifies multi-round assembly designs.
  • To enable the hierarchical assembly of any DNA sequence with high efficiency and precision.

Main Methods:

  • Developed UniClo, a novel DNA assembly method utilizing Type IIs restriction enzyme technology.
  • Employed vector engineering, DNA methylation via recombinant methylases, and CRISPR-dCas9 for steric blockade to regulate methylation.
  • Utilized methylation-sensitive cutting of Type IIs restriction enzyme sites for precise fragment assembly.

Main Results:

  • UniClo enables scarless, one-pot assembly of DNA fragments, even those containing internal recognition sites for the enzyme.
  • The method allows for multi-round hierarchical assembly using a simplified set of three vectors.
  • Assembled plasmids can be recycled as donor plasmids in subsequent assembly rounds, enhancing workflow efficiency.

Conclusions:

  • UniClo provides a simple, universal, and flexible scarless approach for hierarchical DNA assembly.
  • The technology overcomes sequence constraints and unwanted scars associated with traditional methods.
  • UniClo has wide potential applications in synthetic biology, genetic engineering, and molecular cloning.