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Related Concept Videos

PCR01:32

PCR

Overview
Real Time RT-PCR02:57

Real Time RT-PCR

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.
The real-time quantification of the number of amplified products is...
Gene Duplication and Divergence02:37

Gene Duplication and Divergence

The seminal work of Ohno in 1970 popularized the idea of gene duplication and divergence. DNA sequence comparison studies reveal that a large portion of the genes in bacteria, archaebacteria, and eukaryotes was  generated by gene duplication and divergence, indicating its critical role in evolution.
The duplicated copies of the gene are called Paralogs. Paralogs with similar sequences and functions form a gene family. Across several species, a large number of gene families are characterized.

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Shedding new light on stylonychid ciliates: A case study of Tetmemena and Pleurotricha (Ciliophora, Spirotrichea, Hypotrichia, Dorsomarginalia).

European journal of protistology·2026
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Protistan Plankton Responses to Variable Light and Upwelling in the Peruvian Humboldt Current System: Insights Into Community Dynamics Under Environmental Change.

Ecology and evolution·2026
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Bringing the uncultivated microbial majority of freshwater ecosystems into culture.

Nature communications·2025
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Morphological and molecular analyses of season-specific responses of freshwater ciliate communities to top-down and bottom-up experimental manipulations.

mSystems·2025
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Comparing the precision of two digital PCR applications for copy number comparisons in protists.

Scientific reports·2025
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Numerical and Thermal Response of the Bacterivorous Ciliate Colpidium kleini, a Species Potentially at Risk of Extinction by Rising Water Temperatures.

Microbial ecology·2024

Related Experiment Video

Updated: Jul 3, 2026

Determination of Microbial Biomass in Soil using Digital Droplet PCR (ddPCR)
07:46

Determination of Microbial Biomass in Soil using Digital Droplet PCR (ddPCR)

Published on: April 24, 2026

From Gene Copies to Cell Numbers: Advancing Quantitative Approaches in Protistan Ecology Using Digital PCR.

Megan Gross1, Ulrike Koll2, Bettina Sonntag2

  • 1Department of Ecology, Rheinland-Pfälzische Technische Universität Kaiserslautern-Landau, Kaiserslautern, Germany.

Molecular Ecology Resources
|July 2, 2026
PubMed
Summary

Digital PCR (dPCR) offers precise, sensitive quantification of unicellular eukaryotes (protists). Combining dPCR with methods like CARD-FISH advances microbial ecology by improving abundance estimates and understanding protist population dynamics.

Keywords:
CARD‐FISHdPCRenvironmental DNAprotistquantificationrRNA

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Measuring Single-Cell Mitochondrial DNA Copy Number and Heteroplasmy Using Digital Droplet Polymerase Chain Reaction
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Measuring Single-Cell Mitochondrial DNA Copy Number and Heteroplasmy Using Digital Droplet Polymerase Chain Reaction

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Last Updated: Jul 3, 2026

Determination of Microbial Biomass in Soil using Digital Droplet PCR (ddPCR)
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09:15

Measuring Single-Cell Mitochondrial DNA Copy Number and Heteroplasmy Using Digital Droplet Polymerase Chain Reaction

Published on: July 12, 2022

Area of Science:

  • Microbial Ecology
  • Molecular Biology
  • Eukaryotic Microbiology

Background:

  • Quantifying unicellular eukaryote (protist) abundance is crucial for microbial ecology but challenged by methodological variations.
  • High rRNA gene copy numbers in protists complicate accurate molecular quantification.
  • Digital PCR (dPCR) shows promise for absolute quantification, but its application and comparison to existing methods for protists require further investigation.

Purpose of the Study:

  • To develop and validate species-specific digital PCR (dPCR) assays for freshwater ciliates.
  • To establish gene copy number correction factors for accurate dPCR-based abundance estimates.
  • To compare the performance of dPCR with catalyzed reporter deposition-FISH (CARD-FISH) for protist quantification in laboratory and environmental settings.

Main Methods:

  • Development of species-specific dPCR assays for Urotricha castalia and Urotricha pseudofurcata.
  • Determination of gene copy number correction factors for dPCR.
  • Benchmarking dPCR against CARD-FISH using controlled experiments and environmental samples.

Main Results:

  • dPCR assays demonstrated high accuracy and superior precision compared to CARD-FISH under controlled conditions.
  • Method-dependent differences were observed in environmental samples, highlighting both technical and biological variability.
  • dPCR exhibited higher sensitivity, enabling robust detection of low-abundance protist taxa.

Conclusions:

  • Digital PCR, with appropriate correction strategies, provides a scalable and sensitive method for reliable protist quantification.
  • Integrating dPCR with microscopy-based methods like CARD-FISH offers complementary insights into protist population dynamics.
  • This integrative approach advances quantitative microbial ecology and improves abundance estimation for unicellular eukaryotes.