Optimisation_of_high_throughput_16S_rRNA_gene_amplicon_sequencing__an_assessment_of_PCR_pooling__mastermix_use_and_contamination
16S rRNA gene sequencing remains a vital component in characterising human and environmental microbiomes that contribute to health and disease. Sequencing at scale facilitates better powered studies and bacterial surveillance but is limited by cost and time. We identified two areas in our 16S rRNA gene library preparation protocol where modifications had the potential to provide efficiency gains, including (1) pooling of multiple PCR reactions per sample to reduce PCR drift,and (2) manual preparation of mastermix to reduce liquid handling. We studied the impact of these on the alpha and beta diversity, and compositional abundance of nasal samples from healthy human participants and a serially diluted mock microbial community