Long-Read Sequencing and Assembly of the Landsberg Erecta Strain of Arabidopsis with the PacBio® RS II

Significant advances in single-molecule-sequencing read lengths enable highly contiguous genome assemblies for bacteria and model genomes.  Improvements in raw-read error correction, assembly algorithms, and assembly polishing of Pacific Biosciences’ long-read data have led to high-quality assemblies that rival pre-second generation clone-by-clone sequencing efforts.  As a demonstration, we have whole-genome shotgun sequenced the Landsberg erecta (Ler-0)  strain of Arabidopsis thaliana and produced a highly contiguous  assembly in which many full chromosome arms are represented as single contigs.  The contig N50 for the assembly is 6.2 Mbases with the longest contig exceeding 12.5 Mbases.  The average base quality of this assembly is ~Q50.  Library preparation methods including shearing and size selection, sequencing chemistry and assembly methods will be presented.  Additionally, we will present data using the most recent chemistry release (P5-C3).