Metagenomic libraries construction and sequencing

SP Sisi Pu
MW Meng Wang
JW Jinglei Wang
QZ Qian Zhang
XM Xin Ma
RW Rui Wang
SY Sijiu Yu
LW Libin Wang
YP Yangyang Pan
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Extracted genomic DNA was firstly fragmented by sonication to a size of 350 bp, before being end-repaired, A-tailed, and adaptor-ligated using the NEBNext® ΜLtra™ DNA Library Prep Kit for Illumina (NEB, USA) according to the preparation protocol. DNA fragments with lengths of 300–400 bp were amplified by PCR. Finally, PCR products were purified using the AMPure XP system (Beckman Coulter, Brea, CA, USA), and the libraries were analyzed for size distribution using a 2100 Bioanalyzer (Agilent, Santa Clara, CA, USA) and quantified using real-time PCR. Genome sequencing was performed using the Illumina NovaSeq 6000 sequencer with paired-end technology (PE 150) [22].

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