Sorangineae-enriched library of the V3-V4 regions of the 16S rRNA gene
Soil DNA extraction: Fast DNA spin kit for soil (Bio 101 Corp., Vista, CA)   Primer pairs:917F/W6RC   The forward primer contains the 454 Life Sciences primer A sequence (5'-CCATCTCATCCCTGCGTGTCTCCGAC-3'), a four-base linker sequence (GACT), a unique barcode used to tag each PCR product(5'-ACTAGTGATA-3') and the 16S primer(5'-GAATTGACGGGGGCCC-3')  The Reverse primer contains the 454 Life Sciences primer B sequence (5'-CCTATCCCCTGTGTGCCTTGGCAGTC-3'), a four-base linker sequence (GACT), and the 16S primer(5'-GACTCTTATCCGTACTGAGG-3')  Target region:to amplify the V6-V8 region of the 16S rRNA gene sequences  PCR reactions were performed in ten 20 µl reactions with 0.5 µM forward and reverse primers, 50 ng of template DNA, 2×GC buffer II (TAKARA, Japan), 0.2 mM dNTP, certified DNA-free PCR water and 0.5 U of LA Taq (TAKARA, Japan). To further reduce amplification bias, the number of PCR cycles was reduced to 25. Thermal cycling consisted of an initial denaturation at 94°C for 3 minutes followed by 25 cycles of denaturation at 94°C for 30 seconds, annealing at 57°C for 30 seconds, and extension at 72°C for 30 seconds, with a final extension of 10 minutes at 72°C.