PKS 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 Type I polyketide synthase (PKS) ketosynthase/acyl transferase (KS/AT) domains. First-round PCR was performed using degenerate primer set K1F: TSAAGTCSAACATCGGBCA and M6R: CGCAGGTTSCSGTACCAGTA. Second-round PCR incorporated a 5 block and universal primer sequence (UPF-K1F: /5AmMC6/-GCAGTCGAACATGTAGCTGACTCAGGTCAC-K1F and UPR-M6R/5AmMC6/-TGGATCACTTGTGCAAGCATCACATCGTAG-M6R). A third-round PCR was applied to each sample in duplicate to incorporate SMRTcell barcodes for Pacbio RS II sequencing. Run data has been demultiplexed for downstream analysis.