High Throughput Single Molecule DNA Sequencing

描述(申请人提供)：人类基因组计划花了几年时间才完成，但它只是一个时期的开始，在这个时期，医学诊断、法医学和发育生物学需要大量的DNA和RNA序列信息。传统的测序技术在成本、速度和灵敏度方面都有局限性，对序列信息的需求远远超过了目前获得它的能力。我们最近展示了一项新技术的第一个原理实验证明，该技术将为DNA和RNA测序提供一种快速、低成本和高度并行的技术。这项技术在DNA聚合酶将标记的dNTPs加入到固定的单个DNA分子中后，使用单分子检测荧光标记的核苷酸。与目前的测序方法相比，这项技术的主要优势在于它能够并行地从数百万个独立的分子中获得序列信息。在这里，我们建议开发单分子测序的试剂和方法，具有更长的读取长度和更高的准确性，最终达到NIH 99.99%的黄金标准，同时将哺乳动物基因组测序的成本降低到10万美元以下。

DESCRIPTION (provided by applicant): The Human Genome Project took several years to complete, yet it is only the beginning of a period in which large amounts of DNA and RNA sequence information will be required for medical diagnostics, forensics, and developmental biology. Conventional sequencing technology has limitations in cost, speed, and sensitivity and the demand for sequence information far outstrips the current ability to obtain it. We recently demonstrated the first proof of principle experiments for a new technology that will provide a fast, low cost, and highly parallel technique for DNA and RNA sequencing. This technology uses single molecule detection of fluorescently labelled nucleotides after DNA polymerase incorporates labeled dNTPs into immobilized individual DNA molecules. A major advantage of this technique over current sequencing methods lies in its ability to obtain sequence information from millions of independent molecules in parallel. Here we propose to develop reagents and methods for single molecule sequencing runs with longer read lengths and higher accuracy, ultimately reaching the NIH gold standard of 99.99%, while reducing the cost of sequencing a mammalian genome to below $100,000.