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. 2021 Dec 6;11:23456. doi: 10.1038/s41598-021-02837-4

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Specificity evaluation for 25(OH)D₃ detection. (a) Aptamer binding specificity check with different negative controls 25(OH)D₂ and CDA. Maximum fluorescence intensity was observed in 25(OH)D₃ when compared to the other compounds proving the specificity of the aptamer. (b) Quantitative and qualitative analysis was also proved using 3% agarose gel electrophoresis. [Apt – 100 nM, GO-50 µg/mL and 25(OH)D₃-5 µg/mL] F0: only Aptamer which is 1 and F1: Apt + GO in the presence and absence of compounds. The comparison of negatives was expressed as Mean ± S.D., n = 3.