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Nitrogen Cycle Evaluation (NiCE) Chip for Simultaneous Analysis of Multiple N Cycle-Associated Genes
Mamoru Oshiki1, Takahiro Segawa2,3, Satoshi Ishii4
1Department of Civil Engineering, National Institute of Technology, Nagaoka College, Nagaoka, Japan.
Applied and Environmental Microbiology
|February 11, 2018
Summary
A new microfluidic qPCR chip, the nitrogen cycle evaluation (NiCE) chip, enables simultaneous quantification and library preparation for multiple nitrogen cycle genes. This high-throughput tool advances the study of microbial communities involved in nitrogen cycling in environmental samples.
Area of Science:
- Environmental microbiology
- Molecular biology
- Biotechnology
Background:
- Microorganisms drive crucial nitrogen (N) cycle processes.
- Analyzing N cycle functional genes is vital for understanding environmental N transformations.
- Conventional methods for analyzing multiple N cycle genes are often cumbersome and expensive.
Purpose of the Study:
- To introduce and evaluate a novel microfluidic qPCR chip technology for high-throughput analysis of N cycle functional genes.
- To enable simultaneous quantification and amplicon library preparation for multiple N cycle genes and 16S rRNA markers.
- To assess the utility of this technology in environmental samples like wastewater.
Main Methods:
- Development and application of the nitrogen cycle evaluation (NiCE) chip, a microfluidic qPCR platform.
- Simultaneous quantification of multiple N cycle functional genes and 16S rRNA genes.
- Comparison of NiCE chip results with conventional qPCR and amplicon sequencing.
Main Results:
- The NiCE chip provided quantitative results comparable to conventional qPCR.
- The technology was successfully applied to analyze N cycle gene abundance and diversity in wastewater samples.
- Nonspecific amplification was observed but can be addressed through future primer optimization.
Conclusions:
- The NiCE chip offers a high-throughput solution for quantifying and preparing sequence libraries for multiple N cycle functional genes.
- This technology significantly advances the exploration of N cycling in diverse environmental settings.
- The NiCE chip is suitable for analyzing temporal dynamics of N cycle gene transcription, such as in wastewater treatment bioreactors.
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