Since the beginning of the microbiome era decades ago, 16S rRNA gene-based amplicon sequencing has been widely adopted for microbial composition profiling. Many researchers might assume the 16S sequencing workflow is now well-optimized, and selecting a heavily-cited protocol is trivial. However, there are aspects and challenges in 16S sequencing that should be considered before adopting a protocol.
This white paper from Zymo Research outlines challenges in 16S rRNA sequencing to be considered when adopting a library preparation protocol, including selecting a hypervariable region, controlling PCR chimera formation, and normalization, and describes how the Quick-16S Plus NGS Library Prep Kit addresses those challenges.
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