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Related Experiment Video

Updated: Feb 17, 2026

Efficient Nucleic Acid Extraction and 16S rRNA Gene Sequencing for Bacterial Community Characterization
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Sampling, Extraction, and High-Throughput Sequencing Methods for Environmental Microbial and Viral Communities.

Pedro J Torres1, Scott T Kelley2

  • 1Department of Biology, San Diego State University, San Diego, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 11, 2017
PubMed
Summary

High-throughput sequencing advances microbial and viral community studies. Proper experimental design, sample acquisition, and data quality are crucial for valid results in microbiome research.

Keywords:
16S/18S ribosomal RNA gene sequencesDNA extractionMicrobiomeViral purificationViral-like particles (VLPs)

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Area of Science:

  • Microbiology
  • Genomics
  • Bioinformatics

Background:

  • High-throughput sequencing technologies have revolutionized the study of microbial and viral communities.
  • Understanding complex microbial ecosystems is vital, yet many microbes and viruses remain uncultured.
  • High-quality data is essential for reliable insights into these communities.

Purpose of the Study:

  • To outline current methodologies for studying microbial and viral communities.
  • To emphasize the importance of robust experimental design and data quality in high-throughput sequencing studies.
  • To guide researchers in sample acquisition, DNA extraction, library preparation, and sequencing.

Main Methods:

  • Detailed methods for microbial and viral sample acquisition.
  • DNA extraction protocols for diverse environmental samples.
  • Sequencing preparation and execution on Illumina high-throughput platforms.
  • Considerations for experimental design and statistical power.

Main Results:

  • Established protocols for generating high-quality data from microbial and viral communities.
  • Highlighted the critical role of experimental design, including site selection and contamination control.
  • Emphasized the necessity of sufficient sample sizes for statistical validity.

Conclusions:

  • Adherence to rigorous methods ensures the generation of high-quality data for microbiome research.
  • Proper experimental design and adequate sample numbers are as critical as analytical tools.
  • Careful attention to sampling and contamination control is paramount for accurate microbial and viral community analysis.