自体荧光
聚丙烯酰胺
DNA
DNA提取
荧光
链霉亲和素
生物分子
聚合
聚合酶链反应
化学
色谱法
萃取(化学)
材料科学
纳米技术
生物化学
聚合物
生物素
高分子化学
有机化学
物理
基因
量子力学
作者
Jun Wang,Shuyan Zhang,Xinhui Xu,Yujun Xing,Zongru Li,Jinke Wang
出处
期刊:ACS omega
[American Chemical Society]
日期:2020-06-04
卷期号:5 (23): 13829-13839
被引量:5
标识
DOI:10.1021/acsomega.0c01181
摘要
The fast and cost-effective DNA extraction is critical for all DNA-based detections. Here, we fabricated a new kind of polyacrylamide microsphere (PAMMP) in various sizes with two methods, spot polymerization (large size but low yield) and modified inverse microemulsion polymerization (small size but high yield). The fabricated PAMMPs have strong autofluorescence (fPAMMPs), including both visible fluorescence (VF) and near-infrared fluorescence (NIRF), which can remain very stable in various stringent conditions including strong acid and alkali and high temperature. The fabricated fPAMMPs were also highly positively charged, which could be used to effectively capture various biomolecules such as IRDye 800-labeled streptavidin and DNA. We thus developed a new method for rapid extraction (3-5 min) of DNA from various samples including bacteria, mammalian cells, plant and animal solid tissues, and human blood plasma using fPAMMPs. Moreover, the DNA captured on fPAMMPs (fPAMMP@DNA) could be effectively detected by both normal and quantitative PCR amplifications. Finally, we showed that NaBH4 treatment removed autofluorescence in fPAMMPs (PAMMPs), which could also be applied to DNA extraction and PCR detection. In conclusion, we here fabricated new kinds of fPAMMPs and PAMMPs, developed a new rapid DNA extraction method, and demonstrated their useful applications in PCR detection.
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