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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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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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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.
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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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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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Automated Robotic Liquid Handling Assembly of Modular DNA Devices
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[DNA assembly technologies: a review].

Hanchen Chang1,2, Chen Wang1,2, Peixia Wang1,2

  • 1Frontier Science Center for Synthetic Biology, Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, China.

Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology
|December 28, 2019
PubMed
Summary
This summary is machine-generated.

This review covers DNA assembly technologies for synthetic biology, including enzyme-dependent, non-enzymatic, and microbial recombination methods for efficient DNA fragment joining.

Keywords:
DNA assemblyenzyme-dependenthomologous recombinationsynthesissynthetic biology

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Area of Science:

  • Synthetic biology
  • Molecular biology
  • Biotechnology

Background:

  • DNA assembly is fundamental to synthetic biology, enabling the construction of novel biological systems.
  • Advancements have led to diverse DNA assembly methods, including enzyme-dependent and non-enzyme-dependent techniques.
  • Large DNA fragment assembly often relies on microbial recombination for efficiency.

Purpose of the Study:

  • To review and categorize current DNA assembly technologies.
  • To highlight methods suitable for various DNA fragment sizes.
  • To discuss the evolution and applications of DNA assembly in synthetic biology.

Main Methods:

  • Review of enzyme-dependent DNA assembly (e.g., Gibson Assembly, Golden Gate).
  • Analysis of non-enzyme-dependent DNA assembly techniques for automation.
  • Examination of in vivo homologous recombination for large DNA fragment assembly.

Main Results:

  • Enzyme-dependent methods offer precision for smaller constructs.
  • Non-enzymatic methods facilitate high-throughput and automated DNA assembly.
  • Microbial recombination is crucial for assembling kilobase to megabase DNA fragments.

Conclusions:

  • A diverse toolkit of DNA assembly methods exists, catering to different synthetic biology needs.
  • The choice of method depends on fragment size, desired throughput, and automation requirements.
  • Continued innovation in DNA assembly is critical for advancing synthetic biology.