Targeted Sequencing for the Long-Read Era

It is widely known but not often acknowledged that the overwhelming majority of clinical genomics research and practice is performed through some form of targeted sequencing. Furthermore, despite more than a decade of... [ view full abstract ]

It is widely known but not often acknowledged that the overwhelming majority of clinical genomics research and practice is performed through some form of targeted sequencing. Furthermore, despite more than a decade of next-generation sequencing, Sanger sequencing is still the most commonly used clinical sequencing method.  In this talk, I’ll start by discussing some of technical reasons for the delay in penetration of newer sequencing technologies into clinical research.  Then I will discuss how long reads and freedom from systematic error make single molecule, real-time (SMRT®) sequencing a powerful tool for these applications.  I’ll finish with a survey of available targeting methods available for SMRT sequencing, including BAC clones, PCR, target capture as well as a new protocol based on the CRISPR/Cas-9 gene editing system that allows targeting without amplification, thus providing access to regions of the genome previously prone to pseudogene trans-reannealing and polymerase slippage in PCR that destroy crucial genetic information in clinically important loci.