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Molecular Tools in Environmental Microbiology

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Microbiology".

Deadline for manuscript submissions: 31 December 2025 | Viewed by 815

Special Issue Editors


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Guest Editor
Laboratory of Genetics, Institute of Botany, Nature Research Centre, Akademijos Str. 2, LT-08412 Vilnius, Lithuania
Interests: yeasts; killer; microbiota; Sarcocystis; essential oils; microorganisms; viruses; RNA; DNA

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Guest Editor
Laboratory of Genetics, Institute of Botany, Nature Research Centre, Akademijos Str. 2, LT-08412 Vilnius, Lithuania
Interests: transcriptomics; metagenomics; biocontrol yeasts; yeast viruses
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Special Issue Information

Dear Colleagues,

Environmental microbiology continues to face significant technical and analytical challenges. One of the primary obstacles is the extraction of high-quality genomic DNA from complex and inhibitor-rich environmental samples such as soil, feces, sediments, wastewater, and biological fluids. These matrices often contain humic substances, polysaccharides, and other PCR inhibitors that compromise downstream molecular analyses. Another major limitation is the low abundance of target microorganisms within a given environmental sample. This presents difficulties in both the enrichment and specific detection of species of interest, especially in cases where the microbial signal is overwhelmed by a high background of non-target DNA. The advent of next-generation sequencing (NGS) technologies has revolutionized the field, yet several critical limitations remain. These include the scarcity of reference genome sequences for many environmental microbes, the incompleteness and inconsistency of current taxonomic databases, and the challenge of selecting appropriate genetic markers for accurate species-level identification and functional analysis.

The goal of this Special Issue is to present recent advances in molecular methods for studying microorganisms in complex environmental samples. It focuses on overcoming challenges in DNA extraction, PCR inhibition, detection of low-abundance species, and limitations of current sequencing tools and databases. The issue aims to provide practical solutions and highlight innovative tools for accurate microbial identification and functional analysis in environmental microbiology.

We invite microbiologists, molecular ecologists, and environmental scientists to contribute their insights in the following areas and related topics:

  • Experimental Methods and Technical Challenges (improvement methods for DNA/RNA extraction from environmental matrices; strategies to overcome PCR inhibition; novel approaches for microbial enrichment and target-specific amplification; advances in primer design and genetic marker selection; and so on).
  • Sequencing Technologies and Applications (applications of metagenomics, metatranscriptomics, and single-cell sequencing in environmental microbiology).
  • Applications and Case Studies (case studies using molecular tools to monitor microbial communities; standardization and reproducibility in environmental microbiology).
  • Bioinformatics and Database Development (bioinformatics tools and pipelines for environmental microbial analysis; challenges and solutions in microbial reference database development).

Dr. Živilė Strazdaitė-Žielienė
Dr. Juliana Lukša
Guest Editors

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Keywords

  • environmental microbiology
  • molecular methods
  • microbial community analysis
  • low-abundance microorganisms
  • microbial identification
  • environmental DNA
  • next-generation sequencing (NGS)
  • high-throughput sequencing

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Published Papers (1 paper)

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Research

26 pages, 4476 KB  
Article
Benchmarking Cost-Effective DNA Extraction Kits for Diverse Metagenomic Samples
by Andrey Sobolev, Daria Sibiryakina, Elizaveta Chevokina, Darya Slonova, Daria Yurikova, Svetlana Kozlova, Anna Trofimova, Vasiliy Zubarev, Alexander Kiselev, Olga Konovalova, Dmitry Sutormin and Artem Isaev
Int. J. Mol. Sci. 2025, 26(23), 11616; https://doi.org/10.3390/ijms262311616 - 30 Nov 2025
Viewed by 371
Abstract
Extraction of high-quality microbial DNA remains a critical bottleneck in metagenomic research. Environmental samples often produce fragmented DNA and are prone to contaminations that interfere with downstream sequencing, while widely used commercial kits can be prohibitively expensive. Therefore, systematic evaluation of cost-effective alternatives [...] Read more.
Extraction of high-quality microbial DNA remains a critical bottleneck in metagenomic research. Environmental samples often produce fragmented DNA and are prone to contaminations that interfere with downstream sequencing, while widely used commercial kits can be prohibitively expensive. Therefore, systematic evaluation of cost-effective alternatives is essential to support large-scale metagenomic studies. In this work, we benchmarked eight commercial DNA extraction kits from Magen, SkyGen, and Sileks against Qiagen reference kits. Four representative sample types were analyzed: freshwater, seafloor sediments, Pacific oyster (Magallana gigas) gut microbiome, and mammalian feces. DNA yield, integrity, purity, PCR inhibitor content, and eukaryotic DNA admixture were assessed. Microbial community composition, alpha diversity, reproducibility, and contamination (“kitome” and “splashome”) were further evaluated using 16S rRNA amplicon sequencing. We revealed that several alternative kits performed comparably or better than the Qiagen reference standard. Magen Soil and Magen Bacterial provided high yields and reproducibility, though the latter produced more fragmented DNA. SkyGen Stool excelled with host-associated samples, while Sileks Soil and Metagenomic kits preserved higher diversity in sediments. Magen Microbiome consistently underperformed. This study identifies multiple cost-effective DNA extraction strategies and provides practical guidance for selecting balanced DNA purification methods for different sample types. Full article
(This article belongs to the Special Issue Molecular Tools in Environmental Microbiology)
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