16S rRNA for “Granulated straw incorporation with rotary tillage increases the content of soil organic carbon fractions, available nutrients, and shifts bacterial communities in East China”

Bacterial DNA was extractedfrom 0.5 g of fresh soilusing Fast® DNA SPIN Kit (MP Biomedicals). The V3-V4 region of the 16S rRNA gene was amplified with bacterial primers 338F (5′-ACTCCTAGGGAGGAGCA-3′) and 806R (5′-GGACTCHVGGGTWTTAT-3′). The PCR products were purified with the Agarose Gel DNA Purification Kit (TaKaRa). High-through paired-end sequencing of the DNA samples was performed at Majorbio Bio Technology Co. Ltd(Shanghai, China). The raw sequences were analyzed by QIIME software (v 1.9.0). After quality filtering, reads were clustered into operational taxonomic units (OTUs) using the closed-reference OTU picking against the Greengene database in QIIME (gg_13_5 version). Then, the community composition of each sample was assessed at each level. Mothur was used to analyze the αdiversity of the bacterialcommunities (Chao1, Simpson and Shannon indexes).