An inexpensive and simple method for thermally stable immobilization of DNA on an unmodified glass surface: UV linking of poly(T)10-poly(C)10-tagged DNA probes
An inexpensive and simple method for thermally stable immobilization of DNA on an unmodified glass surface: UV linking of poly(T)10-poly(C)10-tagged DNA probes
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DOI：
10.2144/000112905
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发表时间：
2008-09-01
期刊：
影响因子：
2.7
通讯作者：
Wolff, Anders
中科院分区：
文献类型：
--
作者：
Gudnason, Haukur;Dufva, Martin;Wolff, Anders
在载玻片上打印的微阵列通常通过将修饰的寡核苷酸探针共价连接到衍生化表面来构建，其花费相当大。在这篇文章中，我们证明了14个碱基的寡核苷酸与聚（T）10聚（C）10尾（TC标签），但在其他方面未修饰的，可以连接到一个平面，未修饰的玻璃表面的紫外线照射。固定在未改性的玻璃显微镜载玻片上的探针与结合在商业氨基硅烷涂层载玻片上的探针进行类似的操作，并且具有相当的检测限。连接到未改性玻璃的TC标记的探针在再杂交之前在100 ° C的水中孵育20分钟后没有显示出杂交性能的任何显著降低，表明TC标记和未改性玻璃之间的共价键。探针用于热微测序循环反应。此外，TC标签提高了氨基硅烷表面上的固定化探针的杂交性能，表明将TC标签添加到DNA探针的一般益处。总之，我们的研究结果表明，使用TC标记的DNA探针固定在未改性的玻璃表面是一个强大的，热稳定的，非常简单，和廉价的方法制造DNA微阵列。
Microarrays printed on glass slides are often constructed by covalently linking modified oligonucleotide probes to a derivatized surface at considerable expense. In this article, we demonstrate that 14-base oligonucleotides with a poly(T)10 - poly(C)10 tail (TC tag), but otherwise unmodified, can be linked by UV light irradiation onto a plain, unmodified glass surface. Probes immobilized onto unmodified glass microscope slides performed similarly to probes bound to commercial amino-silane-coated slides and had comparable detection limits. The TC-tagged probes linked to unmodified glass did not show any significant decrease in hybridization performance after a 20 min incubation in water at 100 degrees C prior to rehybridization, indicating a covalent bond between the TC tag and unmodified glass. The probes were used in thermal minisequencing cycling reactions. Furthermore, the TC tag improved the hybridization performance of the immobilized probes on the amino-silane surface, indicating a general benefit of adding a TC tag to DNA probes. In conclusion, our results show that using TC-tagged DNA probes immobilized on an unmodified glass surface is a robust, heat-stable, very simple, and inexpensive method for manufacturing DNA microarrays.