DNA微阵列
DNA
杂交探针
核酸
荧光团
核酸热力学
荧光
生物
微阵列
大肠杆菌
化学
核糖核酸
基因
分子生物学
生物化学
基因表达
物理
量子力学
作者
Tomoyuki Taguchi,Machi Ishikawa,Momoko Ichikawa,Takashi Tadenuma,Yuko Hirakawa,Tomoko Yoshino,Yoshiaki Maeda,Hiyori Takeuchi,Daisuke Nojima,Takeo Tanaami,Tadashi Matsunaga
标识
DOI:10.1016/j.bios.2021.113659
摘要
In this study, we developed a novel DNA microarray system that does not require fluorophore-labeling, amplification, or washing of the target nucleic acid fragments. Two types of DNA probes (so-called "signaling probes") labeled with a fluorescence dye (Cy3) and quencher molecule (BHQ2) were spotted on the DNA microarray such that fluorescent signals of Cy3 could be quenched by BHQ2 due to duplex formation between the probes. The addition of the target DNA or RNA fragments disrupted the duplex formed by the probes, resulting in the generation of fluorescence signals. We examined the assay conditions of the signaling probe-based DNA microarray, including the design of the probes, hybridization temperatures, and methods for fragmentation of target molecules. Since this approach does not require time-consuming processes, including labeling, amplification, and washing, the assay achieved specific detection of 16S rDNA and 16S rRNA extracted from Escherichia coli within 60 min, which was significantly rapid compared to conventional PCR-dependent DNA microarrays.
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