FLUORESCENCE CORRELATION SPECTROSCOPY IN CELLS W/ TWO PHOTON EXCITATION

鉴于人类基因组数据的利用， 无先验的单细胞遗传信息测序 扩增将提高基因功能的速度和敏感性 承认。我们建议通过以下方式进行单分子测序 在生产过程中逐步添加碱基对之后 由给定(未知)模板链的互补DNA链 酶DNA聚合酶。将每个基地合并为 聚合酶活性部位是通过检测特定的 荧光标记。赤霉菌DNA合成的初步研究 利用荧光染料PicoGreen的固定化模板链 (分子探针公司)，研究表明，分子的单个分子 可以检测到新合成的DNA。接下来，我们尝试检测 分离PicoGreen分子与新生成的 具有荧光相关光谱设置的DNA。DNA 测序方法也将利用纳米颗粒成分来 DNA操纵和可能的液体流动。

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.