Fast DNA Extraction with Polyacrylamide Microspheres for Polymerase Chain Reaction Detection

文献类型： 外文期刊

第一作者： Wang, Jun

作者： Wang, Jun;Zhang, Shuyan;Xu, Xinhui;Wang, Jinke;Xing, Yujun;Li, Zongru

作者机构：

期刊名称：ACS OMEGA （ 影响因子：3.512； 五年影响因子：3.613 ）

ISSN： 2470-1343

年卷期： 2020 年 5 卷 23 期

页码：

收录情况： SCI

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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