ena-STUDY-ILLUMINA, INC.-13-11-2014-00:52:36:273-1185
DNA Sequencing Using Polymerase Substrate Binding Kinetics
Next-generation sequencing (NGS) has transformed genomic research by decreasing the cost of sequencing.  However, whole genome sequencing is still costly and complex for diagnostics and therapeutics purposes.1, 2  In the clinical space, targeted sequencing has the advantage of allowing researchers to focus on specific genes of interest.3  Routine clinical use of targeted NGS mandates small inexpensive instruments, fast turnaround time, and an integrated and robust workflow.4  Here, we demonstrate a version of the Sequencing by Synthesis (SBS) chemistry that potentially can become a preferred targeted sequencing method in the clinical space.  This sequencing chemistry uses natural nucleotides and is based on real-time recording of the differential polymerase/DNA binding kinetics in the presence of correct or mismatch nucleotides.  This ensemble SBS chemistry has been implemented on existing Illumina sequencing platforms with integrated cluster amplification.  We discuss the advantages of this sequencing chemistry for targeted sequencing as well as its limitations for other genome applications.
Next-generation sequencing (NGS) has transformed genomic research by decreasing the cost of sequencing.  However, whole genome sequencing is still costly and complex for diagnostics and therapeutics purposes.1, 2  In the clinical space, targeted sequencing has the advantage of allowing researchers to focus on specific genes of interest.3  Routine clinical use of targeted NGS mandates small inexpensive instruments, fast turnaround time, and an integrated and robust workflow.4  Here, we demonstrate a version of the Sequencing by Synthesis (SBS) chemistry that potentially can become a preferred targeted sequencing method in the clinical space.  This sequencing chemistry uses natural nucleotides and is based on real-time recording of the differential polymerase/DNA binding kinetics in the presence of correct or mismatch nucleotides.  This ensemble SBS chemistry has been implemented on existing Illumina sequencing platforms with integrated cluster amplification.  We discuss the advantages of this sequencing chemistry for targeted sequencing as well as its limitations for other genome applications.