Sequencing the “Unsequenceable” Single-Molecule, Real-Time (SMRT™) DNA Sequencing: Technology Overview and Recent Applications

Abstract
Understanding the dynamics of biological processes is fundamental to understanding life itself. At Pacific Biosciences, we are developing applications to observe individual biomolecules at work in real time. The first is monitoring DNA synthesis by single DNA polymerase molecules, allowing the speed, processivity, and efficiency of the enzyme to be exploited for new capabilities in DNA sequencing. The power of this new sequencing technology – characterized by long readlengths and fast run times – is highlighted through examples from diverse applications, such as finishing genomes (de novo hybrid assembly), targeted resequencing of difficult to sequence regions like GC- or AT-rich, characterizing transcript and gene fusion diversity, rapid pathogen sequencing, and the direct detection of epigenetic base modifications by making use of the kinetic information collected during DNA-sequencing.