Optimization of high-throughput 16S rRNA gene amplicon sequencing: an assessment of PCR pooling, mastermix use and contamination.

"pmc pmc10634443 pmc10634443 10634443 37843887 37843887 001115 10 1099/mgen 0 001115 methods microbial communities optimization of high-throughput 16s rrna gene amplicon sequencing: an assessment of pcr pooling mastermix use and contamination http://mgen microbiologyresearch org all genomic data generated and/or analysed during the current study are available from the european nucleotide archive (ena) under study accessions prjeb64004 and prjeb64269 *correspondence: dinesh aggarwal da295@cam ac uk + these authors contributed equally to this work 2023 16 10 2023 16 10 2023 [ comment ] 9 10 001115 01 8 2023 28 9 2023 (c) 2023 the authors2023 https://creativecommons org/licenses/by/4 0/ this is an open-access article distributed under the terms of the creative commons attribution license.; we demonstrate no requirement for pooling of pcr amplifications or manual preparation of pcr mastermix resulting in a more efficient 16s rrna gene pcr protocol microbiome nasal samples16s rrna gene laboratory protocol pcr pooling pcr drift mastermix contamination efficiency openaccessembargo 0 all genomic data generated and/or analysed during the current study are available from the european nucleotide archive (ena) under study accessions prjeb64004 and prjeb64269 (see supplementary material 1 available in the online version of this article for additional details)."

"Conflicts of interest The authors declare that there are no conflicts of interest."

"Funding information D.A. is a Clinical PhD Fellow and gratefully supported by the Wellcome Trust (grant no. 222903/Z/21/Z). The CARRIAGE study is funded by Wellcome Collaborative Award in Science (grant no. 211864/Z/18/Z). Work conducted at the Wellcome Sanger Institute was supported by the Wellcome Trust (grant no. 220540/Z/20 /A)."