A Memoir of Inventing Real-Time PCR and Developing the ABI 7700.

Real-time PCR (qPCR) is today's definitive quantitative technology in molecular biology and diagnostics. Until 30 years ago, PCR product analyses were generally performed after amplification using gel-based methods. Quantification typically relied on visual inspection or densitometry of end-point products and was therefore relatively unreliable and poorly suited to high-throughput automation. To celebrate real-time PCR's 30-year anniversary of commercial availability, Professor Stephen Bustin, Guest Editor for the special edition, "Advancing Molecular Science Through Reproducible qPCR: MIQE Guidelines and Beyond," asked Russell Higuchi to give a historical account on how his idea of real-time PCR was conceived and brought to fruition. Dr. Higuchi then asked his collaborator, Lincoln McBride, who drove the development of the ABI 7700-the high-throughput real-time PCR instrument that gave researchers access to this technology-to co-author this dual memoir. This story is told from the perspectives of the two scientists most directly responsible for making real-time PCR practical and widely accessible. Taking turns, Russell Higuchi describes the conceptual and experimental steps at Cetus and then Roche that led from homogeneous PCR detection to continuous fluorescence monitoring, whilst Lincoln McBride details ABI's parallel efforts to commercialize Russ's invention. Together, they trace how experimental insight, engineering constraints, product development, and commercial decision-making shaped the Applied Biosystems 7700 Sequence Detection System and established real-time PCR as a practical and reliable quantitative technology. Their team's efforts persevered through technological uncertainty and within a complex corporate collaboration. They share key historical documents in their original form. Their accounts show how the 7700 system emerged as the convergence of chemistry, optics, software, and product development. The eventual global reliance on real-time PCR during the COVID-19 pandemic demonstrated, at unprecedented scale, the profound and enduring impact of these early technical and organizational choices.