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Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
Published on: August 29, 2014
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Data analysis for 16S microbial profiling from different benchtop sequencing platforms
Victor S Pylro1, Luiz Fernando W Roesch2, Daniel K Morais1
1Microbiology Department, Universidade Federal de Viçosa, Viçosa, MG 36570-900, Brazil; AgroEcology Department, Rothamsted Research, Harpenden, Herts AL52JQ, United Kingdom.
Journal of Microbiological Methods
|September 7, 2014
Summary
Different next-generation sequencing (NGS) technologies yield consistent microbial ecology findings. Applying strict quality control and robust clustering ensures reliable results across diverse sequencing platforms.
Area of Science:
- Microbial Ecology
- Bioinformatics
- Genomics
Background:
- Evaluating microbial ecology studies is challenging due to variations in sequencing methodologies.
- Inconsistent results hinder the comparison of data generated by different next-generation sequencing (NGS) technologies.
Purpose of the Study:
- To investigate whether different NGS technologies can produce comparable results in microbial ecology.
- To identify key factors influencing the consistency of sequencing data.
Main Methods:
- Comparison of data from multiple NGS platforms.
- Application of stringent sequence quality filtering protocols.
- Utilization of accurate clustering algorithms for sequence analysis.
Main Results:
- Consistent biological conclusions were achieved across different NGS technologies.
- Stringent quality filtering and accurate clustering were critical for data comparability.
- Methodological variations did not impede the attainment of similar ecological insights.
Conclusions:
- The choice of NGS technology is less critical than data processing for microbial ecology.
- Standardized bioinformatics pipelines, including quality filtering and clustering, are essential for reproducible microbial ecology research.
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