Pacbio sequencing of 5 Antarctic soil bacterial isolates
Secondary metabolite biosynthetic gene clusters (BGCs) such as NRPS and PKS usually span long, repetitive, high G+C regions difficult to resolve with short-read platforms. Here, long-read sequencing was employed to optimise capture of BGCs. The resolution provided by long reads was also hypothesised to allow for the sequencing of multiple genomes from different genera in one sequencing library. Pooled gDNA was subjected to 10-20 kb BluePippin size selection (Sage Science, Beverly, MA, USA) and a single library prepared using the SMRTbell template preparation kit 1.0 (Pacific Biosciences, Menlo Park, CA) and sequenced on the Pacbio RS II instrument with P6/C4 chemistry, employing three SMRT cells per library.