Sung Kyung Eun, Burns Mark A
Department of Chemical Engineering and Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan 48109-2136, USA.
Anal Chem. 2006 May 1;78(9):2939-47. doi: 10.1021/ac051662f.
We have found that the surface and bulk solution properties in a microfabricated device affect the degree and probability of electrostretching of DNA molecules. Using lambda phage DNA, we found that significantly hydrophilic surfaces between the electrodes decrease the efficiency of stretching. Surfaces treated with higher silane (trimethylchlorosilane) concentrations performed better presumably due to the decreased nonspecific adsorption of DNA on these surfaces compared to their more hydrophilic counterparts. The shape and dimensions of the electrodes also affected the efficiency of stretching. Both liftoff and metal etching methods produced electrodes with random microscopic peaks along the electrode's edge and were poorly suited for stretching. Annealing the electrodes (450 degrees C for 10 min) removed most of these peaks and allowed for more controlled stretching to be obtained. We also found that thin electrodes (65 nm) gave close to a 90% success rate of DNA stretching but stretching with thick electrodes (350 nm) produced only a 20% success rate.
我们发现,微加工设备中的表面和本体溶液特性会影响DNA分子电拉伸的程度和概率。使用λ噬菌体DNA,我们发现电极之间显著亲水的表面会降低拉伸效率。用较高浓度硅烷(三甲基氯硅烷)处理的表面表现更好,这可能是因为与更亲水的表面相比,DNA在这些表面上的非特异性吸附减少。电极的形状和尺寸也会影响拉伸效率。剥离法和金属蚀刻法都会产生沿电极边缘有随机微观峰的电极,不太适合拉伸。对电极进行退火处理(450℃,10分钟)可以去除大部分这些峰,并能获得更可控的拉伸效果。我们还发现,薄电极(65纳米)的DNA拉伸成功率接近90%,而厚电极(350纳米)拉伸的成功率仅为20%。