Donggang bulk soil 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 (5- CCTACGGGNGGCWGCAG-3) and 805R (5-GACTACHVGGGTATCTAATCC-3)which contains Nextera consensus and adaptor sequences at the forward (5- TCGTCGGCAGCGTC-AGATGTGTATAAGAGACAG-target sequence-3) and reverse (5- GTCTCGTGGGCTCGG-AGATGTGTATAAGAGACAG-target sequence-3) (Fadrosh, et al., 2014). The PCR products were cleaned-up and amplified with primers containing Illumina dual indices and sequencing adapters using the forward index i5 (5-AATGATACGGCGACCACCGAGATCTACAC-55555555-TCGTCGGCAGCGTC-3) and reverse i7 (5-CAAGCAGAAGACGGCATACGAGAT-7777777-GTCTCGTGGGCTCGG-3). 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).
Donggang bulk soil wild metagenome rep 1