DNA-seq Cement 11th
DNA extraction was carried out using the NucleoSpin kit following the NucleoSpin Soil Manual protocol. polymerase chain reaction (PCR) amplification was conducted in two stages. The initial PCR targeted the V4 region of the 16S rRNA gene using 515F/806RC primers and KAPA HiFi Hot Start Ready Mix (TaKaRa Bio Inc., Shiga, Japan). Following the denaturation process, a clean-up step was performed using the NucleoSpin Gel and PCR Clean-up Kit to remove free primers and primer dimer species. The second PCR was employed to attach dual indices and Illumina sequencing adapters using the Nextera XT V2 Index Kit.