The challenges of sequencing by synthesis

Area of Science:

• Biochemistry
• Chemistry
• Physics
• Engineering
• Genomics

Background:

• DNA sequencing-by-synthesis (SBS) is a core technology in second-generation DNA sequencing systems.
• Current SBS platforms have achieved significant performance but face challenges in cost reduction for human genome sequencing.

Purpose of the Study:

• To identify and articulate key challenges hindering cost reduction and performance enhancement in DNA sequencing-by-synthesis.
• To solicit input from a broad scientific and engineering community to accelerate advancements in genome sequencing technology.

Main Methods:

• Review of existing DNA sequencing-by-synthesis technologies and their limitations.
• Identification of interdisciplinary challenges spanning sample preparation, surface chemistry, enzyme-substrate optimization, fluorescent labeling, optics, and instrumentation.
• Framing challenges to encourage broad community engagement and input.

Main Results:

• Key challenges include sample preparation, surface chemistry, fluorescent labels, enzyme-substrate system optimization, optics, and instrumentation.
• Tradeoffs between throughput and accuracy, and limitations in read length and phasing require further investigation.
• The need for integrated scientific and technological efforts across multiple disciplines is highlighted.

Conclusions:

• Overcoming current limitations in DNA sequencing-by-synthesis requires a multidisciplinary approach.
• Collaborative efforts are essential to reduce sequencing costs and enhance throughput and quality for widespread genome sequencing.
• Further research and innovation are needed to achieve the goal of sub-$100,000 human genome sequencing.