Chrisey L A, O'Ferrall C E, Spargo B J, Dulcey C S, Calvert J M
Naval Research Laboratory, Washington, DC 20375-5348, USA.
Nucleic Acids Res. 1996 Aug 1;24(15):3040-7. doi: 10.1093/nar/24.15.3040.
Two photolithographic methods are described for the formation of patterned single or multiple DNA species on SiO2 substrates. In the first approach, substrates are treated with a photochemically labile organosilane monolayer film. Irradiation of these surfaces with patterned deep UV (193 nm) light results in patterned chemically reactive groups which are then reacted with heterobifunctional crosslinking molecules. Covalent attachment of modified synthetic DNA oligomers to the crosslinker results in stable DNA patterns. Alternatively, a photoresist is spin-coated over a silane film which had been previously modified with the heterobifunctional crosslinker. Upon patterned irradiation and subsequent development, the underlying crosslinker-modified layer is revealed, and is then reacted with a chemically modified DNA. Feature dimensions to 1 micron are observed when a single fluorescent DNA is attached to the surface. By performing sequential exposures, we have successfully immobilized two distinguishable DNA oligomers on a single surface. Synthetic DNA immobilized in this manner retains the ability to hybridize to its complementary strand, suggesting that these approaches may find utility in the development of miniaturized DNA-based biosensors.
本文描述了两种用于在二氧化硅（SiO₂）衬底上形成图案化的单种或多种DNA分子的光刻方法。在第一种方法中，衬底用具有光化学不稳定性的有机硅烷单层膜进行处理。用图案化的深紫外光（193纳米）照射这些表面会产生图案化的化学反应性基团，然后这些基团与异双功能交联分子发生反应。将修饰的合成DNA寡聚物共价连接到交联剂上可形成稳定的DNA图案。另外，在先前已用异双功能交联剂修饰过的硅烷膜上旋涂光刻胶。经过图案化照射和随后的显影后，露出下面的交联剂修饰层，然后使其与化学修饰的DNA发生反应。当在表面附着单个荧光DNA时，可观察到特征尺寸达到1微米。通过进行连续曝光，我们已成功地在单个表面上固定了两种可区分的DNA寡聚物。以这种方式固定的合成DNA保留了与其互补链杂交的能力，这表明这些方法可能在基于DNA的微型生物传感器的开发中具有实用价值。
