FLUORESCENCE CORRELATION SPECTROSCOPY IN CELLS W/ TWO PHOTON EXCITATION

具有两个光子激发的细胞荧光相关光谱

• 批准号: 6349390
• 负责人: PETRA SCHWILLE
• 金额: $ 0.31万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6349390
• 关键词: bioengineering /biomedical engineering; biological products; biomedical equipment development; biomedical resource; genome; lasers; technology /technique

In view of the utilization of human genome data the capability of sequencing the genetic information of single cells without prior amplification would potentiate speed and sensitivity of gene function recognition. We propose to carry out single molecule sequencing by following the stepwise addition of base pairs during production of the complementary DNA strand of a given (unknown) template strand by the enzyme DNA polymerase. The incorporation of each base at the polymerase active site is to be recognized by detection of a specific fluorescent marker. Preliminary studies on DNA synthesis on immobilized template strands utilized the fluorescent dye PicoGreen (Molecular Probes Inc.), and it is shown that individual molecules of newly synthesized DNA can be detected. Next, we attempt to detect the separate binding events of PicoGreen molecules to the newly generated DNA with a fluorescence correlation spectroscopy setup. The DNA sequencing approach will also utilize nanofabricated compon ents for DNA manipulation and possible fluid flow.

鉴于人类基因组数据的利用 对没有先验的单个细胞的遗传信息进行测序 扩增将增强基因功能的速度和灵敏度 认出。 我们建议通过 在生产过程中逐步添加碱基对 给定（未知）模板链的互补DNA链 酶DNA聚合酶。 每个基部在 聚合酶活性位点应通过检测特定的识别 荧光标记。 关于DNA合成的初步研究 固定的模板链利用了荧光染料picogreen （分子探针Inc.），显示出单个分子的分子 可以检测到新合成的DNA。 接下来，我们尝试检测 PICOGREEN分子的单独结合事件与新生成的 具有荧光相关光谱设置的DNA。 DNA 测序方法还将利用纳米制造的组件 DNA操纵和可能的流体流动。