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High Throughput Sequencing Using Single Molecule Millikan Sequencing

使用单分子密立根测序进行高通量测序

基本信息

批准号：
8215614
负责人：
Javier Farinas
金额：
$ 17.78万
依托单位：
CAERUS MOLECULAR DIAGNOSTICS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-01-26 至 2013-11-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): The feasibility of single molecule Millikan Sequencing for de novo sequencing of mammalian genomes for under $1,000 will be evaluated. This novel sequencing-by-synthesis approach measures the increased charge as nucleotides are added to a single DNA template attached to a tethered bead without any need for labels. Opposing electrical, hydrodynamic and restoring forces result in bead motion that is a function of the length of DNA attached to the bead. Simultaneous optical detection of an array of millions of beads undergoing chain elongation will allow high-throughput sequencing. Model calculations and preliminary results indicate that this method should enable accurate, long read length and label-free DNA sequencing. The lack of labels leads to negligible reagent costs while the relatively simple dark-field optics leads to a low-cost instrument. Long read lengths will result in low genome assembly cost. The single template approach eliminates the need for emulsion or bridge PCR, making sample preparation much easier and cheaper. The aim of the proposed two-year exploratory project is to demonstrate the feasibility of sequencing a single DNA template by monitoring changes in the intrinsic charge of a growing DNA chain. PUBLIC HEALTH RELEVANCE: Nucleic acid sequence information is critical to medical research and to basic biological studies. The goal of this project is to demonstrate the feasibility of DNA sequencing system based on a label-free detection approach, single molecule Millikan Sequencing, which may be capable of de novo sequencing of mammalian genomes for under $1,000. Such a platform will allow researchers and clinicians to perform the translational research ultimately required to improve human health.

描述（由申请人提供）：将评估用于哺乳动物基因组从头测序的单分子密立根测序的可行性，费用低于1,000美元。这种新的合成测序方法测量了增加的电荷，因为核苷酸被添加到连接到拴系珠的单个DNA模板中，而不需要任何标记。相反的电、流体动力和恢复力导致珠运动，其是附着于珠的DNA长度的函数。同时光学检测数百万个经历链延伸的珠粒的阵列将允许高通量测序。模型计算和初步结果表明，这种方法应该能够实现准确的，长的读取长度和无标记的DNA测序。标签的缺乏导致试剂成本可以忽略不计，而相对简单的暗场光学器件导致低成本仪器。长读段长度将导致低基因组组装成本。单模板方法消除了对乳液或桥式PCR的需要，使样品制备更容易和更便宜。拟议的两年期探索项目的目的是通过监测不断增长的DNA链的内在电荷变化来证明对单个DNA模板进行测序的可行性。公共卫生相关性：核酸序列信息对医学研究和基础生物学研究至关重要。该项目的目标是证明基于无标记检测方法的DNA测序系统的可行性，即单分子密立根测序，该系统可能能够以低于1000美元的价格对哺乳动物基因组进行从头测序。这样一个平台将使研究人员和临床医生能够进行最终改善人类健康所需的转化研究。