16S amplicon sequencing of river sediment metagenome: Rainfall Runoff Impact Stage of channel D
Total genomic DNA was extracted from river sediment samples using DNeasy PowerSoil Kit (Qiagen). The V4-V5 hypervariable region of the 16S rRNA gene was amplified with primers 515FGTGCCAGCMGCCGCGGTAA and 806R: GGACTACNNGGGTATCTAAT. PCR amplification was performed under the following conditions: 95C for 3 min, followed by 30 cycles of 95C for 30 s, 55C for 30 s, 72C for 45 s, and a final extension at 72C for 10 min. The PCR products were purified and used to construct sequencing libraries with Nextera XT DNA Library Preparation Kit (Illumina). Libraries were sequenced on an Illumina MiSeq platform using 2 300 bp paired-end sequencing.