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Development and Evaluation of Whole-Cell and Genomic DNA-Based Next-Generation Sequencing Standards

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"Development and Evaluation of Whole-Cell and Genomic DNA-Based Next-Generation Sequencing Standards"

The increasing accessibility of next-generation sequencing (NGS) technologies has greatly influenced microbiome analyses, with applications in clinical, diagnostic, therapeutic, industrial, and environmental research. However, the complexity of 16S rRNA and metagenomic sequencing analysis presents significant challenges due to biases introduced during sample preparation, DNA extraction, PCR amplification, library preparation, sequencing, or data interpretation.

A primary challenge in assay standardization is the limited availability of reference materials. To address these biases and provide a measure of standardization within microbiome research and applications, ATCC has developed a set of mock microbial communities, which includes lyophilized whole cells or genomic DNA, for use as NGS Standards in microbiome research. These standards mimic mixed metagenomics samples and comprise fully sequenced, characterized strains selected based on select phenotypic and genotypic attributes. In collaboration with ATCC, One Codex has developed a data analysis module that provides simple output in the form of true-positive, relative abundance, and false-negative scores for 16S rRNA community profiling and shotgun metagenomic sequencing.

This white paper from ATCC discusses the development and evaluation of the NGS standards for use in various applications, such as method optimization and data interpretation.

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