PCR Bias Impacts Microbiome Ecological Analyses.

ABSTRACT: Polymerase Chain Reaction (PCR) is a critical step in amplicon-based microbial community profiling, allowing the selective amplification of marker genes such as 16S rRNA from environmental or host-associated samples. Despite its widespread use, PCR is known to introduce amplification bias, where some DNA sequences are preferentially amplified over others due to factors such as primer-template mismatches, sequence GC content, and secondary structures. Although these biases are known to affect transcript abundance, their implications for ecological metrics remain poorly understood. In this study, we conduct a comprehensive evaluation of how PCR-bias influences both within-samples ( α -diversity) and between-sample ( β -diversity) analyses. We show that perturbation-invariant di

SUBMITTER: Rathod D 

PROVIDER: S-EPMC12324424 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature