title: Fungal community in rhizosphere of Gastrodia elata at Young stage
design: To generate fungal libraries, the first round of PCR was performed to amplify the ITS region using primers ITS3-F (5-GCA TCG ATG AAG AAC GCA GC-3) and ITS4-R (5- TCC TCC GCT TAT TGA TAT GC-3) which contains Nextera consensus and adaptor sequences at the forward and reverse. The PCR products were cleaned-up and amplified with primers containing Illumina dual indices and sequencing adapters using the forward index i5 and reverse i7. The PCR conditions were as follows: initial denaturation at 95 C for 3min, followed by 25 cycles of denaturation at 95 C for 30s, annealing at 55 C for 30s, extension at 72 C for 30s, and final extension at 72 C for 5min. Purified amplicon libraries were pooled and sequenced at CJ bioscience Inc. (South Korea) with an Illumina MiSeq system using the MiSeq Reagent Kit v2 (Illumina Inc., USA