High-resolution nucleic acid detection using online polyacrylamide gel electrophoresis platform

Zihao Wang; Yiren Cao; Zixian Yu; Youli Tian; Jicun Ren; Weiwen Liu; Liuyin Fan; Qiang Zhang; Chengxi Cao

doi:10.1016/j.chroma.2023.464571

High-resolution nucleic acid detection using online polyacrylamide gel electrophoresis platform

J Chromatogr A. 2024 Jan 4:1713:464571. doi: 10.1016/j.chroma.2023.464571. Epub 2023 Dec 9.

Authors

Zihao Wang¹, Yiren Cao², Zixian Yu¹, Youli Tian³, Jicun Ren⁴, Weiwen Liu¹, Liuyin Fan⁵, Qiang Zhang⁶, Chengxi Cao⁷

Affiliations

¹ School of Sensing Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China.
² School of Sensing Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China; School of Chemistry and Chemical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China.
³ School of Sensing Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China; School of Life Science and Biotechnology, State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai, 200240, China.
⁴ School of Chemistry and Chemical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China.
⁵ Student Innovation Center, Shanghai Jiao Tong University, Shanghai, 200240, China. Electronic address: lyfan@sjtu.edu.cn.
⁶ School of Sensing Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China. Electronic address: billy_zq@sjtu.edu.cn.
⁷ School of Sensing Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China; School of Life Science and Biotechnology, State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai, 200240, China. Electronic address: cxcao@sjtu.edu.cn.

PMID: 38091846
DOI: 10.1016/j.chroma.2023.464571

Abstract

Polyacrylamide gel electrophoresis (PAGE) is one of the most popular techniques for the separation and detection of nucleic acids. However, it requires a complicated detection procedure and offline detection format, which inevitably leads to band broadening and thus compromises the separation resolution. To overcome this problem, we developed an online PAGE (OPAGE) platform by integrating the gel electrophoresis apparatus with the gel imaging system, so as to obviate the need for the complicated detection procedure. Notably, OPAGE enabled the real-time monitoring of the separation process and the immediate imaging of the separation results once the electrophoresis ended. Using a series of synthetic DNAs with different lengths as samples, we demonstrated that the OPAGE platform enhanced 32-64 % of the number of theoretical plates, showed a robust dynamic range of 0.1-12.5 ng/μL, and realized a limit of detection as low as 0.08 ng/μL DNA. Based on our results, we anticipate that the OPAGE platform is a promising alternative to traditional nucleic acid gel electrophoresis for simple and high-resolution detection and quantification and nucleic acid.

Keywords: High-resolution separation; Nucleic acid; Online detection; Polyacrylamide gel electrophoresis.

MeSH terms

DNA*
Electrophoresis, Polyacrylamide Gel
Nucleic Acids*

Substances

DNA
Nucleic Acids