title: Amplicon_seq of soil metagenome: soil
design: PCR reactions were performed in a 25 ŚĚL mixture containing 1.0 ŚĚL of each primer at 10 ŚĚM, 2.0 ŚĚL of template DNA (20 ng/ŚĚL), 10ĄÁ buffer 2.5 ŚĚL, 0.125 ŚĚL Pyrobest DNA Polymerase (5U/ŚĚL, Takara), 2.0 ŚĚL of dNTP (deoxy-ribonucleoside triphosphate) at 2.5 mM and 16.375 ŚĚL of ultrapure sterile water. The following thermal program was used for amplification: 94ĄăC for 4 min, followed by 27 cycles of denaturation at 94ĄăC for 30 s, annealing at 55ĄăC for 45 s