Betanzos-Cabrera Gabriel, Harker Brent W, Doktycz Mitchel J, Weber James L, Beattie Kenneth L
Area Académica de Nutrición, Instituto de Ciencias de la Salud, Universidad Autónoma del Estado de Hidalgo, Abasolo 600 Pachuca de Soto, Hidalgo, 42000, Mexico.
Mol Biotechnol. 2008 Jan;38(1):71-80. doi: 10.1007/s12033-007-9001-z. Epub 2007 Sep 6.
Two different solid supports, channel glass and flat glass, were compared for their affect on the sensitivity and efficiency of DNA hybridization reactions. Both solid supports were tested using a set of arrayed, synthetic oligonucleotides that are designed to detect short insertion/deletion polymorphisms (SIDPs). A total of 13 different human SIDPs were chosen for analysis. Capture probes, designed for this test set, were covalently immobilized on substrates. Hybridization efficiency was assessed using fluorescently labeled stacking probes which were preannealed to the target and then hybridized to the support-bound oligonucleotide array; the hybridization pattern was detected by fluorescence imaging. It was found that structural features of nucleic acid capture probes tethered to a solid support and the molecular basis of their interaction with targets in solution have direct implications on the hybridization process. Our results demonstrate that channel glass has a number of practical advantages over flat glass including higher sensitivity and a faster hybridization rate.
比较了两种不同的固体支持物——通道玻璃和平板玻璃，它们对DNA杂交反应的灵敏度和效率的影响。使用一组排列好的合成寡核苷酸对这两种固体支持物进行测试，这些寡核苷酸旨在检测短插入/缺失多态性（SIDP）。总共选择了13种不同的人类SIDP进行分析。为该测试集设计的捕获探针被共价固定在底物上。使用荧光标记的堆积探针评估杂交效率，这些探针预先与靶标退火，然后与固定在支持物上的寡核苷酸阵列杂交；通过荧光成像检测杂交模式。发现固定在固体支持物上的核酸捕获探针的结构特征及其与溶液中靶标的相互作用分子基础对杂交过程有直接影响。我们的结果表明，通道玻璃比平板玻璃具有许多实际优势，包括更高的灵敏度和更快的杂交速率。
