Journal
NATURE BIOTECHNOLOGY
Volume 34, Issue 9, Pages 942-+Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nbt.3601
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Funding
- Minnesota Partnership for Biotechnology and Medical Genomics (grant MNP IF) [14.09]
- Margot Marsh Biodiversity Foundation
- US National Institutes of Health (PharmacoNeuroImmunology Fellowship NIH/NIDA) [T32 DA007097-32]
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Amplicon-based marker gene surveys form the basis of most microbiome and other microbial community studies. Such PCR-based methods have multiple steps, each of which is susceptible to error and bias. Variance in results has also arisen through the use of multiple methods of next-generation sequencing (NGS) amplicon library preparation. Here we formally characterized errors and biases by comparing different methods of amplicon-based NGS library preparation. Using mock community standards, we analyzed the amplification process to reveal insights into sources of experimental error and bias in amplicon-based microbial community and microbiome experiments. We present a method that improves on the current best practices and enables the detection of taxonomic groups that often go undetected with existing methods.
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