DNA sequencing using polymerase substrate-binding kinetics

Michael John Robert Previte, Chunhong Zhou, Matthew Kellinger, Rigo Pantoja, Cheng-Yao Chen, Jin Shi, BeiBei Wang, Amirali Kia, Sergey Etchin, John Vieceli, Ali Nikoomanzar, Erin Bomati, Christian Gloeckner, Mostafa Ronaghi and Molly Min He ()
Additional contact information
Michael John Robert Previte: Protein Engineering, Illumina Inc.
Chunhong Zhou: Engineering, Illumina Inc.
Matthew Kellinger: Protein Engineering, Illumina Inc.
Rigo Pantoja: Protein Engineering, Illumina Inc.
Cheng-Yao Chen: Protein Engineering, Illumina Inc.
Jin Shi: Protein Engineering, Illumina Inc.
BeiBei Wang: Protein Engineering, Illumina Inc.
Amirali Kia: Protein Engineering, Illumina Inc.
Sergey Etchin: Engineering, Illumina Inc.
John Vieceli: Bioinformatics, Illumina Inc.
Ali Nikoomanzar: Protein Engineering, Illumina Inc.
Erin Bomati: Protein Engineering, Illumina Inc.
Christian Gloeckner: Protein Engineering, Illumina Inc.
Mostafa Ronaghi: Protein Engineering, Illumina Inc.
Molly Min He: Protein Engineering, Illumina Inc.

Nature Communications, 2015, vol. 6, issue 1, 1-12

Abstract: Abstract 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 purposes. In the clinical space, targeted sequencing has the advantage of allowing researchers to focus on specific genes of interest. Routine clinical use of targeted NGS mandates inexpensive instruments, fast turnaround time and an integrated and robust workflow. 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 an existing Illumina sequencing platform with integrated cluster amplification. We discuss the advantages of this sequencing chemistry for targeted sequencing as well as its limitations for other applications.

Date: 2015
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DOI: 10.1038/ncomms6936

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