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Immobilization of DNA for Single Molecule Detection

用于单分子检测的 DNA 固定化

基本信息

批准号：
09304070
负责人：
SHIMOMURA Masatsugu
金额：
$ 26.82万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09304070/
关键词：
DNA single molecule fluorescence life time intercalation base sequencing SNOM polyion complex single molecule detection

项目摘要

Double-helical DNA is a supramolecular architecture composed of complementary base-pairings of adenine-thymine and cytosine-guanine based on specific hydrogen bonding, and carries genetic information. Due to the stacking interaction of p-electrons at the ground states of the base-pairs, DNA can act as a p-electron medium for the photoinduced electron transfer. Toward the single molecule detection of DNA by using a novel optical technique based on the combination of near field optics and photoinduced electron transfer, polynucleic acids have been immobilized as two-dimensional molecular assemblies or single molecule by means of the specific intermolecular interaction at the air-water interface. DNA, having phosphate groups in its backbone, was assembled with counter charged amphiphiles and fixed as a polyion complex monolayer at the air-water interface. We have investigated the effect of DNA on the fluorescence lifetime of a binary component monolayer of an acridine amphiphile and a cat … More ionic double chain amphiphile. Strong fluorescence quenching was found when polyG polyC was dissolved in a water subphase. Static quenching of acridine fluorescence, probably due to the electron transfer from guanine base, is occurred because the fluorescence life-time is not so reduced as fluorescence intensity. The photoinduced electron transfer to an ITO electrode was measured by an electrochemical method. Anodic photosensitized current of the amphiphilic acridine dye is enhanced by the complex formation with polynucleic acids. PolyG polyC, whose guanine base is assumed to be *idized by the excited state of the acridine moiety, is most effective for photocurrent generation. By using the simple casting method we have prepared mesoscopic stripe pattern of DNA on a mica substrate. An aqueous DNA solution mixed with/without an aqueous alginic acid solution was dropped onto a fleshly cleaved mica surface and dried by heating up to ca. 80ﾟC.The mesoscopic lines parallel to the receding direction of the solvent were observed in the cast film. When alginic acid were added in the aqueous DNA solution, more regularly aligned stripe patterns were obtained. Height of those lines was below 10 nm. Less

双螺旋DNA是由腺嘌呤-胸腺嘧啶和胞嘧啶-鸟嘌呤碱基互补配对组成的超分子结构，携带遗传信息。由于p电子在碱基对基态的堆积相互作用，DNA可以充当光诱导电子转移的p电子介质。为了利用近场光学和光诱导电子转移相结合的新光学技术对DNA进行单分子检测，利用分子间的特异性相互作用将多核酸固定为二维分子组装体或单分子。DNA，在其骨干中具有磷酸基团，组装与带相反电荷的两亲分子和固定作为一个聚离子复合物单层在空气-水界面。研究了DNA对吖啶两亲分子和猫二元单分子膜荧光寿命的影响 ...更多信息离子双链两亲物。当polyG polyC溶解在水亚相中时，发现强烈的荧光猝灭。吖啶荧光的静态猝灭可能是由于鸟嘌呤碱基的电子转移，因为荧光寿命没有像荧光强度那样降低。通过电化学方法测量到ITO电极的光诱导电子转移。两亲性吖啶染料的阳极光敏化电流通过与多核酸形成复合物而增强。聚G聚C，其鸟嘌呤碱基被假定为通过吖啶部分的激发态被 * 化，对于光电流产生是最有效的。用简单的浇铸法在云母基底上制备了DNA的介观条纹图案。将与/不与海藻酸水溶液混合的DNA水溶液滴到肉裂的云母表面上，并通过加热至约100 ℃干燥。在流延膜中观察到平行于溶剂的后退方向的介观线。当在DNA水溶液中加入海藻酸时，获得了更规则排列的条纹图案。这些线的高度低于10 nm。少