PacBio sequencing holds promise for addressing large-genome complexities, such as long, highly repetitive, low-complexity regions and duplication events that are difficult to resolve with short-read technologies. Several strategies, with varying outcomes, are available for de novo sequencing and assembling of larger genomes. Using a diploid fungal genome, estimated to be ~80 Mb in size, as the basis dataset for comparison, we highlight assembly options when using only PacBio sequencing or a combined strategy leveraging data sets from multiple sequencing technologies. Data generated from SMRT Sequencing was subjected to assembly using different large-genome assemblers, and comparisons of the results will be shown. These include results generated with HGAP, Celera Assembler, MIRA, PBJelly, and other assembly tools currently in development. Improvements observed include a near 50% reduction in the number of contigs coupled with at least a doubling of contig N50 size in genome assemblies incorporating SMRT Sequencing data. We further show how incorporating long reads also highlights new challenges and missed insights of short-read assemblies arising from heterozygosity inherent in multiploid genomes.
Organization: PacBio
Year: 2014
