Conventional wisdom holds that PCR amplification for sequencing should employ pooled replicate reactions to reduce bias due to jackpot effects and chimera formation. However, modern amplicon data analysis employs methods that may be less sensitive to such artifacts. Here we directly compare results from single versus triplicate reactions for 16S amplicon sequencing and find no significant impact of adopting a less labor-intensive single-reaction protocol.

中文翻译：

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16S rRNA 基因扩增子测序不需要进行三次 PCR 反应。

传统观点认为，用于测序的 PCR 扩增应采用合并复制反应，以减少由于大奖效应和嵌合体形成而导致的偏差。然而，现代扩增子数据分析采用的方法可能对此类伪影不太敏感。在这里，我们直接比较 16S 扩增子测序的单次反应与三次重复反应的结果，发现采用劳动强度较低的单反应方案没有显着影响。