DNA extraction, library preparation and sequencing

Binfeng Yu; Lini Jin; Zhouwei Chen; Wanyun Nie; Liangliang Chen; Yanhong Ma; Huan Chen; Yawen Wu; Yunting Ma; Jianghua Chen; Fei Han

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DNA extraction, library preparation and sequencing

BY Binfeng Yu

LJ Lini Jin

ZC Zhouwei Chen

WN Wanyun Nie

LC Liangliang Chen

YM Yanhong Ma

HC Huan Chen

YW Yawen Wu

YM Yunting Ma

JC Jianghua Chen

FH Fei Han

This method is extracted from research article: Ann Transl Med, Aug 2021

The gut microbiome in microscopic polyangiitis with kidney involvement: common and unique alterations, clinical association and values for disease diagnosis and outcome prediction

DOI: 10.21037/atm-21-1315

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Total genomic DNA was extracted from all fecal samples using the PowerSoil^® DNA Isolation Kit (MO BIO, USA) following the manufacturer’s instructions. PCR was then performed to amplify the V3-V4 variable region of 16S rRNA gene. The 16S amplicon library was constructed using the TruSeq^® DNA PCR-Free Sample Preparation Kit (Illumina, USA) and sequenced on an Ion S5TM platform at Zhejiang Institute of Microbiology (Hangzhou, China).

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