title: The challenges of sequencing by synthesis.
abstract: DNA sequencing-by-synthesis (SBS) technology, using a polymerase or ligase enzyme as its core biochemistry, has already been incorporated in several second-generation DNA sequencing systems with significant performance. Notwithstanding the substantial success of these SBS platforms, challenges continue to limit the ability to reduce the cost of sequencing a human genome to $100,000 or less. Achieving dramatically reduced cost with enhanced throughput and quality will require the seamless integration of scientific and technological effort across disciplines within biochemistry, chemistry, physics and engineering. The challenges include sample preparation, surface chemistry, fluorescent labels, optimizing the enzyme-substrate system, optics, instrumentation, understanding tradeoffs of throughput versus accuracy, and read-length/phasing limitations. By framing these challenges in a manner accessible to a broad community of scientists and engineers, we hope to solicit input from the broader research community on means of accelerating the advancement of genome sequencing technology.
authors.affiliation: GE Healthcare Life Sciences, Piscataway, New Jersey, USA. carl.fuller@alumni.upenn.edu
authors.forename: Carl W
authors.initials: CW
authors.lastname: Fuller
authors.forename: Lyle R
authors.initials: LR
authors.lastname: Middendorf
authors.forename: Steven A
authors.initials: SA
authors.lastname: Benner
authors.forename: George M
authors.initials: GM
authors.lastname: Church
authors.forename: Timothy
authors.initials: T
authors.lastname: Harris
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Huang
authors.forename: Stevan B
authors.initials: SB
authors.lastname: Jovanovich
authors.forename: John R
authors.initials: JR
authors.lastname: Nelson
authors.forename: Jeffery A
authors.initials: JA
authors.lastname: Schloss
authors.forename: David C
authors.initials: DC
authors.lastname: Schwartz
authors.forename: Dmitri V
authors.initials: DV
authors.lastname: Vezenov
mesh_terms: D000818:Animals; D004247:DNA / Q000096:biosynthesis*; D005456:Fluorescent Dyes; D006801:Humans; D055620:Optical Phenomena; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods*; D013379:Substrate Specificity; D013499:Surface Properties
chemical_list: D005456:Fluorescent Dyes; D004247:DNA