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Related Concept Videos

DNA Isolation01:24

DNA Isolation

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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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Guided Protocol for Fecal Microbial Characterization by 16S rRNA-Amplicon Sequencing
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A simplified protocol for DNA extraction from FTA cards for faecal microbiome studies.

Amanda Bolt Botnen1, Mads Bjørn Bjørnsen1, Antton Alberdi1

  • 1Center for Evolutionary Hologenomics, The GLOBE Institute, University of Copenhagen, Denmark.

Heliyon
|January 26, 2023
PubMed
Summary

FTA cards offer a simplified DNA extraction method for animal fecal microbiomes. This approach yields higher microbial diversity and an expanded core microbiome compared to traditional freezing and extraction methods.

Keywords:
16S rRNACatsDry preservativesLongitudinal studiesMetabarcodingMetagenomicsMicrobiotaPowerSoil

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Area of Science:

  • Microbiology
  • Genomics
  • Bioinformatics

Background:

  • Metagenomic studies increasingly require data from remote or longitudinal samples.
  • Standard sample collection, storage, and DNA extraction methods are sensitive to variations in time, storage conditions, and chemical processes.
  • FTA cards have been identified as a cost-effective method for preserving animal fecal microbiomes.

Purpose of the Study:

  • To develop and validate a simplified DNA extraction methodology for fecal samples stored on FTA cards.
  • To compare the performance of this simplified FTA card method against a conventional approach (immediate freezing followed by Qiagen PowerSoil DNA isolation kit).

Main Methods:

  • Faecal samples were stored on FTA cards.
  • A simplified DNA extraction protocol was applied to FTA card-stored samples.
  • Microbiome DNA was extracted using the Qiagen PowerSoil DNA isolation kit from immediately frozen faecal samples for comparison.

Main Results:

  • The simplified FTA card DNA extraction methodology resulted in microbial community data with higher diversity.
  • The FTA card method revealed an expanded core microbiome compared to the PowerSoil methodology.
  • The results indicate that the FTA card method is a robust alternative for microbiome analysis.

Conclusions:

  • The simplified FTA card DNA extraction method is a viable and effective alternative for metagenomic studies involving fecal material.
  • This method is particularly useful when traditional freeze-based storage is not feasible.
  • The FTA card approach enhances microbial community analysis by providing higher diversity and a broader core microbiome.