16S rRNA amplification sequencing of sediment: Zostera japonica rhizosphere
The bacterial 16S rRNA gene was amplified using 338F and 806R primers. The PCR reaction mixture (20 l) contained 2 Pro Taq (10 l), forward and reverse primers (4 M), and template DNA (200 ng). PCR cycling conditions were as follows: 1 cycle of 3 min at 95C, 30 cycles of 30 s for denaturation at 95C, 30 s for annealing at 53C and 45 s for elongation at 72C, 1 cycle of 10 min at 72C. The amplified PCR products were purified using an AxyPrep DNA Gel Extraction Kit (Axygen Biosciences, CA, USA) and quantified using QuantiFluor-ST (Promega, USA)