Lawrence Berkeley National Laboratory

The assembly and analysis of microbial species on earth remains a largely unexplored area of life. This is partially due to their inability to be cultured but also based on the large historic cost of drafting and finishing individual microbial species genomes. The single-molecule real-time (SMRTTM) sequencing platform developed by Pacific Biosciences (PacBio) offers several benefits including Single Molecule real-time analysis, longer read length at fast speed, low sequencing redundancy and bias. Thus, it was used at JGI as a quick-turnaround and cost-effective solution for finishing microbial genomes. Construction of PacBio library by traditional protocol still requires micrograms of genomic DNA. In many cases, getting high quantity of genomic DNA remains as a major challenge. Recently, PacBio developed a more efficient library construction method using terminal deoxynucleotidyl transferase (TdT), which makes it possible to obtain sufficient sequencing data for assembly from significantly smaller amount of genomic DNA. We have tested and validated this newly developed method. Preliminary analysis results suggested that this technology can be used for microbial genome assembly with PacBio only data