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DNA Bacteriophages01:26

DNA Bacteriophages

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Bacteriophages, or phages, are viruses that specifically infect bacteria, utilizing their genetic material to hijack host cellular machinery for replication. DNA bacteriophages employ single-stranded DNA (ssDNA) or double-stranded DNA (dsDNA) genomes. These phages exhibit diverse replication strategies and host interactions, influencing their ecological roles and applications in biotechnology and medicine.ssDNA BacteriophagesssDNA phages, with their small genomes, utilize unique strategies to...
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In contrast to the lytic cycle, phages infecting bacteria via the lysogenic cycle do not immediately kill their host cell. Instead, they combine their genome with the host genome, allowing the bacteria to replicate the phage DNA along with the bacterial genome. The incorporated copy of the phage genome is called the prophage. Some prophages can re-activate and enter the lytic cycle. This often occurs in response to a perturbation, such as DNA damage, but can also transpire in the absence of...
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Bacteriophages, also known as phages, are specialized viruses that infect bacteria. A key characteristic of phages is their distinctive “head-tail” morphology. A phage begins the infection process (i.e., lytic cycle) by attaching to the outside of a bacterial cell. Attachment is accomplished via proteins in the phage tail that bind to specific receptor proteins on the outer surface of the bacterium. The tail injects the phage’s DNA genome into the bacterial cytoplasm. In the...
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Metal ions can be separated from one another by complexation with organic ligands–the chelating agent– to form uncharged chelates. Here, the chelating agent must contain hydrophobic groups and behave as a weak acid, losing a proton to bind with the metal. Since most organic ligands used in this process are insoluble or undergo oxidation in the aqueous phase, the chelating agent is initially added to the organic phase and extracted into the aqueous phase. The metal-ligand complex is...
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Filtration Isolation of Nucleic Acids: A Simple and Rapid DNA Extraction Method
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A Rapid Bacteriophage DNA Extraction Method.

Džiuginta Jakočiūnė1, Arshnee Moodley2

  • 1Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg C, Denmark. djak@sund.ku.dk.

Methods and Protocols
|June 6, 2019
PubMed
Summary
This summary is machine-generated.

Researchers developed a rapid DNA extraction method for bacteriophages (phages). This technique simplifies obtaining high-quality phage DNA for whole genome sequencing, aiding antibiotic resistance research.

Keywords:
DNA extractionfastgenomephagesequencingspin column

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Area of Science:

  • Microbiology and Virology
  • Genomics and Molecular Biology

Background:

  • Bacteriophages (phages) are explored as alternatives to antibiotics for combating antibiotic-resistant bacteria.
  • Whole genome sequencing of phages is crucial for understanding their biology and identifying potential virulence factors.
  • Existing phage DNA extraction methods are often time-consuming and require specialized equipment like ultracentrifuges.

Purpose of the Study:

  • To develop a rapid and straightforward method for extracting DNA from double-stranded DNA (dsDNA) bacteriophages.
  • To assess the suitability of the extracted phage DNA for downstream genomic applications.

Main Methods:

  • A novel, simplified DNA extraction protocol was established for dsDNA phages.
  • The method was optimized for phages with titers exceeding 1.0 × 1010 plaque-forming units (PFU)/mL.
  • The entire extraction process was designed to be completed in under four hours.

Main Results:

  • High-quality dsDNA was successfully extracted from bacteriophages using the new method.
  • The extracted phage DNA was compatible with standard library preparation kits, specifically Nextera XT.
  • The DNA yielded sufficient quality for successful sequencing on the Illumina MiSeq platform.

Conclusions:

  • The described method offers a quick and efficient alternative to traditional phage DNA extraction techniques.
  • This simplified protocol facilitates whole genome sequencing of bacteriophages, supporting research into antibiotic resistance and phage therapy.
  • The method's compatibility with common library preparation and sequencing platforms enhances its utility in molecular biology labs.