BACseq: Cost-effective, High-throughput Strategy for BAC-by-BAC Sequencing

Prior to the Next Generation Sequencing (NGS) revolution a typical genome project involved generating BAC libraries, assembling the resulting BACs into minimum tiling paths (MTPs) and the sequencing individual BACs.  Subsequent to the NGS revolution genomes whole genome shotgun (WGS) sequencing has largely replaced BAC-based genome sequencing. The ability to quickly and cost-effectively to generate genome assemblies via the WGS strategy BAC-based approaches still provide the highest quality results, but are typically cost-prohibitive. Although various strategies for pooling BACs prior to sequencing with NGS technologies have been developed, these also suffer from potential repeat-induced artifacts in genome assembly.  In response to the need for cost-effective methods to generate high quality genome sequences, we developed the “BAC-seq” technology, which combines the cost efficiency and high-throughput of NGS with the accuracy of BAC-by-BAC sequencing strategies.  The protocol for BAC-Seq will be described and results of pilot studies presented.