Sorokin N V, Yurasov D Y, Cherepanov A I, Kozhekbaeva J M, Chechetkin V R, Gra O A, Livshits M A, Nasedkina T V, Zasedatelev A S
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov str. 32, Moscow, Russia 119991.
J Biomol Struct Dyn. 2007 Jun;24(6):571-8. doi: 10.1080/07391102.2007.10507146.
Using hydrogel-based oligonucleotide microchips developed previously for the choice of drugs during leukemia treatment and the other diseases, it is shown that the acceleration of external transport by mixing buffer solution with peristaltic pump not only enhances the observable fluorescence signals, but also improves significantly the discrimination between perfect and mismatch duplexes at the intermediate stage of hybridization on the oligonucleotide microchips. The discrimination efficiency for a given hybridization time grows monotonously with the frequency of flow pulsations. The mixing with frequency 10 Hz accelerates the hybridization rate approximately thrice and improves the discrimination efficiency 1.5-2.5 times higher for overnight hybridization. To study these effects, we have developed the special peristaltic pump mixing solution in a hybridization chamber of 35 mul in volume (area approximately 1 x 1 cm(2) and height 0.3 mm). We present also the brief theoretical summary for the interpretation and assessment of the observed experimental features.
使用先前开发的基于水凝胶的寡核苷酸微芯片来选择白血病治疗及其他疾病的药物，结果表明，通过蠕动泵将缓冲溶液混合来加速外部传输，不仅增强了可观测的荧光信号，还显著提高了寡核苷酸微芯片杂交中间阶段完美双链体与错配双链体之间的区分度。在给定的杂交时间下，区分效率随流动脉动频率单调增加。频率为10 Hz的混合使杂交速率加快约三倍，并使过夜杂交的区分效率提高1.5至2.5倍。为研究这些效应，我们开发了一种特殊的蠕动泵，用于在体积为35 μl的杂交室（面积约1×1 cm²，高度0.3 mm）中混合溶液。我们还给出了简要的理论总结，用于解释和评估所观察到的实验特征。
