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Switchable inhibitory behavior of divalent magnesium ion in DNA hybridization-based gene quantification
- Title
- Switchable inhibitory behavior of divalent magnesium ion in DNA hybridization-based gene quantification
- Authors
- Jin, Hyowon; Lim, Hyun Jeong; Liles, Mark R.; Chua, Beelee; Son, Ahjeong
- Ewha Authors
- 손아정
- SCOPUS Author ID
- 손아정
- Issue Date
- 2022
- Journal Title
- ANALYST
- ISSN
- 0003-2654
1364-5528
- Citation
- ANALYST vol. 147, no. 21, pp. 4845 - 4856
- Publisher
- ROYAL SOC CHEMISTRY
- Indexed
- SCIE; SCOPUS
- Document Type
- Article
- Abstract
- Contrary to the understanding that divalent cations only result in under-estimation of gene quantification via DNA hybridization-based assays, we have discovered that Mg2+ could cause either under or over-estimation at different concentrations. Its switchable inhibitory behavior is likely due to its rigid first solvation (hydrated) shell and hence it is inclined to form non-direct binding with DNA. At low concentrations, it caused under-estimation by occupying the hybridization sites. At high concentrations, it caused probe, signaling and target DNA to aggregate non-specifically via Coulomb forces. By quantifying target DNAs at a range of Mg2+ concentrations using a gene quantification assay (NanoGene assay), a Mg2+ inflection concentration of similar to 10(-3) M was observed for both target ssDNA and dsDNA. Field emission scanning electron microscopy (FE-SEM), energy dispersive X-ray spectroscopy (EDS), and Fourier transform infrared spectroscopy (FT-IR) were employed to observe Mg2+-induced non-specific binding in the complexes that mimicked the presence of target DNA. Together with two other divalent cations Ca2+ and Cu2+, they were further examined via zeta potential measurements as well as NanoGene assay. This study revealed the importance of Mg2+ in achieving accurate gene quantification. Through a better mechanistic understanding of this phenomenon, it will be possible to develop strategies to mitigate the impact of Mg2+ on DNA hybridization-based gene quantification.
- DOI
- 10.1039/d2an01164f
- Appears in Collections:
- 공과대학 > 환경공학과 > Journal papers
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