NRPS amplicon by Pacbio RS II: polar soil metagenomes
Extreme environments are attractive targets for microbial biomining due to their potential as reservoirs for novel secondary metabolites. We surveyed > 200 soil samples, comprising nine Antarctic and three High Arctic sites, for non-ribosomal peptide synthetase (NRPS) adenylation (AD) domains. First-round PCR employed degenerate primer sets with additional 5 block and universal primer tags: UPF-A3F: /5AmMC6/-GCAGTCGAACATGTAGCTGACTCAGGTCACGCSTACSYSATSTACACSTCSGG and UPR-A7R: /5AmMC6/-TGGATCACTTGTGCAAGCATCACATCGTAGSASGTCVCCSGTSCGGTAS. A second round PCR incorporated SMRTcell barcodes for Pacbio RS II sequencing. Prior to barcoding, NRPS PCR products were pooled in equimolar amounts representing the start (0, 2 m), middle (100, 102 m) and end (200, 202 m) of each transect for all positive sites. Run data has been demultiplexed for downstream analysis.