EAGER: BISH: Isothermal solid-phase PCR using electrostatic control of DNA hybridization

EAGER：BISH：使用静电控制 DNA 杂交的等温固相 PCR

• 批准号: 0939048
• 负责人: Arnold Vainrub
• 金额: $ 12万
• 依托单位: Auburn University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0939048&HistoricalAwards=false
• 关键词: EAGER; BISH; Isothermal; solid; phase

0939048VainrubThe proposed research investigates the hitherto unknown fundamental characteristics of electrostatic DNA hybridization. Namely; the yield, kinetic rates, ability to denature long DNA and to support a sufficient number of hybridization cycles, in order to establish conditions for efficient PCR amplification. The specific aims are to: 1. Construct a fluorescent detector for hybridization and extension of electrode tethered primers.An evanescent wave fluorescent real-time detector will be built to measure the kinetics and yieldof denaturation, annealing, and extension stages in solid-phase PCR.2. Prove-in-principle and optimize electrostatic isothermal PCR. Linear and exponentialamplification will be demonstrated in real-time PCR mode using oligonucleotide templates.Efficient electrostatic pulse cycles and optimal surface and solution conditions will be defined.Amplification protocols will be developed.

0939048Vainrub拟议的研究调查了迄今为止未知的静电DNA杂交的基本特征。即：产率、动力学速率、变性长DNA的能力和支持足够数量的杂交循环的能力，以建立有效PCR扩增的条件。具体目标是：1.构建用于电极系链引物杂交和延伸的荧光检测器，建立用于固相PCR中变性、退火和延伸阶段的动力学和产量的倏逝波荧光实时检测器.原理证明并优化静电等温PCR。将使用寡核苷酸模板在实时PCR模式中证明线性和指数扩增。将定义有效的静电脉冲周期和最佳的表面和溶液条件。将开发扩增方案。