Donggang root wild
To generate bacterial libraries, the first round of PCR was performed to amplify the V3-V4 region of the 16S rRNA gene using universal primers 341F and 805R 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 94 C for 3min, followed by 25 cycles of denaturation at 94 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 ChunLab Inc. (South Korea) with an Illumina MiSeq system using the MiSeq Reagent Kit v2 (Illumina Inc., USA).
SUB9312821
Donggang root wild metagenome rep 3