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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
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Updated: Jan 12, 2026

Field Identification of Matricaria chamomilla using a Portable qPCR System
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Going Mobile: Using Portable Genomic Technologies for PCR-Free In Situ Species Identification and Real-Time Molecular

Evan J Kipp1, Marissa S Milstein1,2, Lexi E Frank1

  • 1Department of Veterinary and Biomedical Sciences, College of Veterinary Medicine University of Minnesota St. Paul Minnesota USA.

Ecology and Evolution
|November 7, 2025
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Summary

Portable nanopore sequencing with adaptive sampling enables rapid, in-situ genomic analysis for biodiversity monitoring. This method bypasses PCR, streamlining field-based molecular species identification for mammals and insects.

Keywords:
ChiropteraCulicidaePhlebotominaeadaptive samplingmitogenomephylogenetic capture

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Area of Science:

  • Genomics
  • Molecular Biology
  • Ecology

Background:

  • Anthropogenic environmental changes threaten biodiversity and increase zoonotic disease risk via host shifts.
  • Accurate biodiversity assessments and molecular species monitoring are crucial for managing these threats.

Purpose of the Study:

  • To demonstrate the utility of portable laboratories and targeted long-read nanopore sequencing for in-situ genomic and systematic analyses.
  • To assess the effectiveness of nanopore adaptive sampling (NAS) for selective mitochondrial DNA sequencing in field conditions.

Main Methods:

  • Field DNA extraction and library preparation from small mammals and blood-feeding insects in Guyana.
  • Targeted sequencing using nanopore adaptive sampling (NAS) with related mitogenome assemblies as enrichment targets.
  • In-situ mitogenome assembly and generation of mitochondrial biomarker consensus sequences.

Main Results:

  • Complete mitogenomes and consensus biomarker sequences were generated for nine small mammals and four insect species.
  • Molecular identifications were confirmed using local BLAST and maximum likelihood analyses.
  • The NAS approach proved amplification-free, bypassing PCR and streamlining field workflows.

Conclusions:

  • Targeted sequencing with NAS is an effective tool for portable laboratories to enhance field biodiversity monitoring.
  • This method facilitates rapid molecular species assessments in vertebrates and invertebrates, crucial for tracking emerging pathogens.