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Related Concept Videos

DNA Isolation01:24

DNA Isolation

35.3K
DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
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Related Experiment Video

Updated: Apr 29, 2026

Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants
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A universal DNA extraction and PCR amplification method for fungal rDNA sequence-based identification.

A M Romanelli1, J Fu, M L Herrera

  • 1Department of Pathology and Laboratory Medicine, UC Davis Medical Center, Sacramento, CA, USA.

Mycoses
|May 29, 2014
PubMed
Summary

This study presents a rapid, inexpensive DNA extraction protocol for fungal pathogens. The method efficiently yields pure DNA suitable for polymerase chain reaction (PCR) and sequencing across major fungal phyla.

Keywords:
18S RNA28S ribosomal DNAD1/D2ITSdiagnostic

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Area of Science:

  • Mycology
  • Molecular Biology
  • Biotechnology

Background:

  • Accurate identification of fungal pathogens is crucial for clinical diagnostics and research.
  • Sequence-based identification methods, such as polymerase chain reaction (PCR), rely on high-quality template DNA.
  • Existing DNA extraction protocols can be time-consuming, expensive, or require harsh chemicals, limiting their widespread application.

Purpose of the Study:

  • To develop and standardize a rapid, inexpensive DNA extraction protocol for fungal pathogens.
  • To ensure the protocol yields DNA of sufficient purity and quantity for PCR and sequencing.
  • To validate the protocol's applicability across major fungal phyla, including yeasts and filamentous fungi.

Main Methods:

  • A novel DNA extraction protocol combining physical and chemical lysis was developed.
  • The protocol was tested on 519 clinical and culture collection strains of fungi.
  • DNA suitability for PCR was assessed by targeting the ITS and D1/D2 regions for amplification and sequencing.

Main Results:

  • The developed protocol successfully extracted template DNA from all tested fungal strains, representing major phyla.
  • All extracted DNA templates were suitable for PCR amplification and subsequent sequencing.
  • The protocol was completed in approximately 30 minutes, without using organic solvents or ethanol precipitation.
  • The method minimized tube manipulations, enhancing ease of use.

Conclusions:

  • This rapid and inexpensive DNA extraction protocol provides high-quality template DNA suitable for PCR and sequencing.
  • The protocol is broadly applicable to diverse fungal species, including yeasts and filamentous fungi.
  • Its efficiency and simplicity make it a valuable tool for fungal pathogen identification and molecular research.