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Related Experiment Video

Updated: Feb 26, 2026

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
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Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons

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Quantitative metagenomics using a portable protocol.

Kaiqin Bian1, Andrea Busch2, John Norton2

  • 1School of Civil and Environmental Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA.

Applied and Environmental Microbiology
|February 24, 2026
PubMed
Summary
This summary is machine-generated.

A new field-deployable workflow enables rapid, quantitative microbial community profiling using Nanopore sequencing. This approach provides accurate, real-time data for water sector applications, improving process control and wastewater surveillance.

Keywords:
Nanopore sequencingabsolute quantitationinternal standardslimit of quantitationportable laboratorytaxonomic identification

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Last Updated: Feb 26, 2026

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Area of Science:

  • Environmental Microbiology
  • Metagenomics
  • Molecular Diagnostics

Background:

  • Current microbial community profiling methods are often time-consuming and require complex lab equipment.
  • Rapid, quantitative onsite analysis is crucial for timely decision-making in the water sector.
  • Existing portable sequencing technologies often lack quantitative capabilities.

Purpose of the Study:

  • To develop a field-deployable workflow for rapid detection and absolute quantitation (rD+rQ) of microbial communities.
  • To enable quantitative metagenomics using real-time Nanopore sequencing.
  • To provide accurate microbial profiling for environmental and water-related applications.

Main Methods:

  • Integrated high-molecular-weight DNA recovery protocol for diverse environmental samples.
  • Utilized multiplexed Nanopore sequencing with barcoded spike-in-based calibration (BSINC).
  • Established dynamic detection and quantitation limits based on genome coverage and copy number variation.

Main Results:

  • The rD+rQ workflow achieved species-level identification and absolute quantitation comparable to digital PCR.
  • BSINC demonstrated superior calibration accuracy over conventional spike-in methods.
  • The workflow proved effective on complex environmental samples, providing precise microbial data.

Conclusions:

  • The developed rD+rQ workflow offers a portable and user-friendly solution for rapid microbial community analysis.
  • This approach facilitates real-time monitoring and decision-making in the water industry.
  • Enables accurate quantitative data for applications ranging from bioprocess control to wastewater surveillance.