University of Freiburg, Center for Biological Systems Analysis (ZBSA), Habsburgerstrasse. 49, Freiburg, 79104, Germany.
University of Freiburg, Department of Microsystems Engineering (IMTEK), Georges-Köhler-Allee 103, Freiburg, 79110, Germany.
Sci Rep. 2020 Apr 1;10(1):5770. doi: 10.1038/s41598-020-62404-1.
In this work we show how DNA microarrays can be produced batch wise on standard microscope slides in a fast, easy, reliable and cost-efficient way. Contrary to classical microarray generation, the microarrays are generated via digital solid phase PCR. We have developed a cavity-chip system made of a PDMS/aluminum composite which allows such a solid phase PCR in a scalable and easy to handle manner. For the proof of concept, a DNA pool composed of two different DNA species was used to show that digital PCR is possible in our chips. In addition, we demonstrate that DNA microarray generation can be realized with different laboratory equipment (slide cycler, manually in water baths and with an automated cartridge system). We generated multiple microarrays and analyzed over 13,000 different monoclonal DNA spots to show that there is no significant difference between the used equipment. To show the scalability of our system we also varied the size and number of the cavities located in the array region up to more than 30,000 cavities with a volume of less than 60 pL per cavity. With this method, we present a revolutionary tool for novel DNA microarrays. Together with new established label-free measurement systems, our technology has the potential to give DNA microarray applications a new boost.
在这项工作中，我们展示了如何以快速、简便、可靠且具有成本效益的方式在标准显微镜载玻片上批量生产 DNA 微阵列。与传统的微阵列生成方法不同，微阵列是通过数字固相 PCR 生成的。我们开发了一种由 PDMS/铝复合材料制成的腔芯片系统，允许以可扩展且易于处理的方式进行固相 PCR。为了验证概念，我们使用由两种不同 DNA 物种组成的 DNA 池来证明我们的芯片中可以进行数字 PCR。此外，我们还证明了可以使用不同的实验室设备（滑架式循环仪、手动水浴和自动化盒式系统）来实现 DNA 微阵列的生成。我们生成了多个微阵列，并分析了超过 13000 个不同的单克隆 DNA 点，以表明使用的设备之间没有显着差异。为了展示我们系统的可扩展性，我们还将位于阵列区域的腔室的大小和数量变化了 30000 多个，每个腔室的体积小于 60 皮升。通过这种方法，我们为新型 DNA 微阵列提供了一种革命性的工具。结合新建立的无标记测量系统，我们的技术有可能为 DNA 微阵列应用带来新的推动。
