In situ DNA synthesis on glass substrate for microarray fabrication using self-focusing acoustic transducer
In situ DNA synthesis on glass substrate for microarray fabrication using self-focusing acoustic transducer
复制标题
使用自聚焦声换能器在玻璃基板上原位 DNA 合成用于微阵列制造
DOI：
10.1109/tase.2006.871483
复制
发表时间：
2006
影响因子：
5.6
通讯作者：
E. S. Kim
中科院分区：
文献类型：
--
作者：
J. Kwon;S. Kamal;E. S. Kim
本文提出了一种基于液滴喷射的技术，用于在不同的基底上合成脱氧核糖核酸（DNA）序列，如玻璃，塑料或硅。任何DNA序列都可以通过不需要任何喷嘴的自聚焦声换能器（SFAT）喷射DNA碱基的液滴来合成。SFAT可以喷射直径约为3-5 /spl mu/m的液滴，这明显小于商业喷墨打印机喷射的液滴，并减少了合成所需的试剂量。将SFAT阵列与微通道和储存器整合，用于将DNA碱基递送至SFAT。将聚-1-赖氨酸涂覆的载玻片图案化，并用作原位合成多个T碱基的靶基底。与现有的一些商业解决方案相比，这种方案的显著优势在于，它可以让遗传学家在自己的实验室中以负担得起的成本使用计算机程序在几个小时内合成任何DNA序列。本文介绍了按需DNA合成的概念和方案（与声喷射器集成微流控组件）沿着的实际DNA合成的结果由SFAT。脱氧核糖核酸（DNA）微阵列允许遗传学家在芯片中同时监测数千个基因之间的相互作用。有生产DNA微阵列的商业系统，但没有一个能灵活地在遗传学家自己的实验室中按需合成DNA微阵列。Affyoung的基因芯片技术生产DNA探针序列，这些序列是在Affyoung用一组4 n个n-mer光掩模预制的。其他技术通过喷墨打印或接触分配将预制的DNA序列转移到基底（玻璃、塑料或硅）上。安捷伦和罗塞塔使用他们的喷墨打印技术在他们的工厂生产DNA探针序列。用于打印微阵列的喷墨打印头使用压电或热致动，并通过喷嘴喷射液滴。因此，从这些装置喷射的最小液滴尺寸取决于喷嘴的尺寸。小的喷嘴难以构造成具有良好的均匀性并且易于堵塞。本文提出的想法是开发一种基于微机电系统（MEMS）的便携式系统，用于在不同的基底上合成DNA，使用无风扇，无热，无透镜，声学液滴喷射器。未来的研究是使用定向液滴喷射器合成具有不同碱基组合的更长DNA序列。
This paper presents a droplet-ejection-based technique for synthesizing deoxyribonucleic acid (DNA) sequences on different substrates, such as glass, plastic, or silicon. Any DNA sequence can be synthesized by ejecting droplets of DNA bases by a self-focusing acoustic transducer (SFAT) that does not require any nozzles. An SFAT can eject liquid droplets around 3-5 /spl mu/m in diameter, which is significantly smaller than those ejected by commercial ink jet printers and reduces the amount of reagents needed for the synthesis. An array of SFATs is integrated with microchannels and reservoirs for delivery of DNA bases to the SFATs. Poly-l-lysine-coated glass slide is patterned, and is used as a target substrate for in situ synthesis of multiple T bases. The significant advantage of this scheme over some of the existing commercial solutions is that it can allow geneticists to synthesize any DNA sequence within hours using a computer program at an affordable cost in their own labs. This paper describes the concept and scheme of the on-demand DNA synthesis (with an acoustic ejector integrated with microfluidic components) along with the results of an actual DNA synthesis by an SFAT. Note to Practitioners-Deoxyribonucleic acid (DNA) microarrays allow geneticists to monitor the interactions among thousands of genes simultaneously in a chip. There are commercial systems for producing DNA microarrays, but none of them give flexibility to synthesize DNA microarrays on-demand in the geneticist's own lab. Affymetrix's GeneChip technology produces DNA probe sequences premade at Affymetrix with a set of 4n photomasks for n-mers. Other techniques transfer premade DNA sequences to a substrate (glass, plastic, or silicon) through ink-jet printing or contact dispensing. Agilent and Rosetta use their ink-jet printing technology to produce DNA probe sequences at their factories. The ink-jet print heads used for printing microarrays use either piezoelectric or thermal actuation, and eject liquid droplets through nozzles. Thus, the smallest droplet size ejected from these devices depends on the size of the nozzle. The small nozzles are difficult to construct with good uniformity and tend to get clogged. The idea presented in this paper is to develop a microelectromechanical-system (MEMS)-based portable system for synthesizing DNA on different substrates, using nozzleless, heatless, lensless, acoustic droplet ejectors. The future research is to synthesize longer DNA sequences with a combination of different bases, using directional droplet ejectors.