Abstract Basal Stem Rot and Upper Stem Rot diseases caused by pathogenic fungus Ganoderma boninense continue to be a major plight in the palm oil industry. Despite continuous research in combating the problem, resolution remains stagnant. Here, developed an automated, high-throughput DNA extraction protocol on microfluidics device for a quick, disposable, label-free detection, within 2 h of assessment. Microfluidics was designed using AutoCAD software, fabricated on microscopic glass substrate using negative photoresist (SU-8 2015) and molded with a biopolymer silicone, Polydimethylsiloxane. G. boninense and unknown pathogenic fungus isolated from rotten mushroom were grown and fractions of extracted DNA were pooled and analyzed for comparison along with synthetic ssDNA of G. boninense. Results from LPM and HPM show successful fabrication with ≤0.1 mm variance between the dimensions in the design before and after lithography process. The PDMS microfluidics show no leakage when run with DNA samples. Analyses from I-V measurement, UV–vis, FTIR, and PCR show comparable results between extracted and synthetic ssDNA of G. boninense and a contrast with the unknown pathogenic fungus, indicating a successful DNA extraction protocol via microfluidics for label-free identification of G. boninense. Optimization of DNA extraction can be further devised for applicability on lab-on-a-chip devices.

中文翻译：

---

自动化、高通量 DNA 提取方案，适用于油棕病原体 Ganoderma Boninense 的一次性无标签微流体集成 DNA 生物传感器

摘要 由病原菌灵芝引起的基部腐烂病和上茎腐烂病一直是棕榈油行业的一大难题。尽管在解决这个问题方面不断进行研究，但解决方案仍然停滞不前。在这里，在微流体设备上开发了一种自动化、高通量的 DNA 提取协议，可在评估后 2 小时内进行快速、一次性、无标记检测。微流体是使用 AutoCAD 软件设计的，使用负光刻胶 (SU-8 2015) 在微观玻璃基板上制造，并用生物聚合物硅树脂聚二甲基硅氧烷模制。从腐烂的蘑菇中分离出 G. boninense 和未知的致病真菌，并将提取的 DNA 部分汇集起来，并与 G. boninense 的合成 ssDNA 进行比较分析。LPM 和 HPM 的结果表明制造成功，≤0。光刻工艺前后设计尺寸之间的差异为 1 毫米。使用 DNA 样本运行时，PDMS 微流体显示无泄漏。IV 测量、UV-vis、FTIR 和 PCR 的分析显示了 G. boninense 的提取和合成 ssDNA 之间的可比结果以及与未知病原真菌的对比，表明通过微流体技术成功的 DNA 提取方案可用于 G. boninense 的无标记鉴定。博宁。可以进一步设计 DNA 提取的优化以适用于芯片实验室设备。boninense 和与未知致病真菌的对比，表明通过微流体技术成功提取 DNA 协议，用于无标记识别 G. boninense。可以进一步设计 DNA 提取的优化以适用于芯片实验室设备。boninense 和与未知致病真菌的对比，表明通过微流体技术成功提取 DNA 协议，用于无标记识别 G. boninense。可以进一步设计 DNA 提取的优化以适用于芯片实验室设备。