Order340Sample077_16S bacterial community
A second PCR reaction was performed to add Illumina sequencing adapters and sample-specific dual indexes using PCRBIO HiFi (PCR Biosystems Ltd., UK) for 15 cycles. The second PCR products were purified with HighPrep PCR Clean Up System, as described for the first PCR. Sample concentrations were normalized using the SequalPrep Normalization Plate (96) Kit (Thermofisher, USA), following the manufacturer's instructions.