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Raw sequences were processed with Quantitative Insight into Microbial Ecology 2 (QIIME) [24] pipeline using the divisive amplicon denoising algorithm 2 (DADA2) to filter, trim, and merge paired-end reads into amplicon sequence variants (ASVs)

There is no published evidence this is more effective than a single route, but it is a common pattern in community protocols

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: Molecular mechanism of hypoxiamediated hepatic gluconeogenesis by transcriptional regulation