Agencourt a mpure xp nucleic acid purification kit

Total DNA was extracted using the Omega Stool DNA Kit (MoBio Laboratories, Carlsbad, CA, USA). The DNA quality and concentration were measured using spectrophotometry. Using soil DNA as template, upstream primer 338 (5 ′ -ACTCCTACGGGAGGCAGCAG-3 ′ ) and downstream primer 806R (5 ′ -GGACTACNNGGGTATCTAAT-3 ′ ) were used to amplify the V3-V4 region of bacterial 16Sr RNA gene. An 8 bp barcode sequence was added to each of the 5 ′ ends of the upstream and downstream primers to distinguish between different samples. PCR products were detected using 1% Agarose gel electrophoresis and were purified using an Agencourt AMPure XP nucleic acid purification kit (Beckman Coulter, Bria, CA, USA). PCR products were used to construct the microbial diversity sequencing library, the Illumina MiSeq PE300 (Illumina Inc., San Diego, CA, USA) high-throughput sequencing platform was used for paired-end sequencing, and the original sequencing sequences were uploaded to the NCBI SRA database.