Grassland soil Fusarium
Amplicon libraries were created from DNAs isolated from soil using two rounds of PCR. The first round used RPB2-5F2 (5’- GGGGWGAYCAGAAGAAGGC-3’), RPB2-7cR (5’- CCCATRGCTTGYTTRCCCAT-3’). The first round used RPB2-5F2 and RPB2-RP1 (5'-GGNGTCATGCARATCATNGC-3') primers. The first round of PCR used an initial denaturing step at 95°C (5min) followed by 24 cycles of 95°C (1min), 60°C (1min), 72°C (1min), and ended with an extension of 72°C (10min). The second round of PCR used primers amended with 454 GS FLX+ specific adapters and sample-specific barcodes, with an annealing temperature of 65°C. Samples were gel extracted prior to sequencing from the RPB2-5F2 primer.