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Article

The Identification of the Banana Endogenous Reference Gene MaSPS1 and the Construction of Qualitative and Quantitative PCR Detection Methods

1
College of Life Sciences, South China Agricultural University, Guangzhou 510642, China
2
College of Agriculture & Biology, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China
3
Institute of Fruit Tree Research, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Genes 2023, 14(12), 2116; https://doi.org/10.3390/genes14122116
Submission received: 29 October 2023 / Revised: 18 November 2023 / Accepted: 22 November 2023 / Published: 23 November 2023
(This article belongs to the Special Issue Genetically Modified Organisms (GMO) Biosafety)

Abstract

Endogenous reference genes play a crucial role in the qualitative and quantitative PCR detection of genetically modified crops. Currently, there are no systematic studies on the banana endogenous reference gene. In this study, the MaSPS1 gene was identified as a candidate gene through bioinformatics analysis. The conservation of this gene in different genotypes of banana was tested using PCR, and its specificity in various crops and fruits was also examined. Southern blot analysis showed that there is only one copy of MaSPS1 in banana. The limit of detection (LOD) test showed that the LOD of the conventional PCR method is approximately 20 copies. The real-time fluorescence quantitative PCR (qPCR) method also exhibited high specificity, with a LOD of approximately 10 copies. The standard curve of the qPCR method met the quantitative requirements, with a limit of quantification (LOQ) of 1.14 × 10−2 ng—about 20 copies. Also, the qPCR method demonstrated good repeatability and stability. Hence, the above results indicate that the detection method established in this study has strong specificity, a low detection limit, and good stability. It provides a reliable qualitative and quantitative detection system for banana.
Keywords: banana (Musa paradisiaca); endogenous reference gene; MaSPS1; qPCR banana (Musa paradisiaca); endogenous reference gene; MaSPS1; qPCR

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MDPI and ACS Style

Zhu, L.; Lin, Y.; Yang, W.; Pan, Z.; Chen, W.; Yao, J.; Sheng, O.; Zhou, L.; Jiang, D. The Identification of the Banana Endogenous Reference Gene MaSPS1 and the Construction of Qualitative and Quantitative PCR Detection Methods. Genes 2023, 14, 2116. https://doi.org/10.3390/genes14122116

AMA Style

Zhu L, Lin Y, Yang W, Pan Z, Chen W, Yao J, Sheng O, Zhou L, Jiang D. The Identification of the Banana Endogenous Reference Gene MaSPS1 and the Construction of Qualitative and Quantitative PCR Detection Methods. Genes. 2023; 14(12):2116. https://doi.org/10.3390/genes14122116

Chicago/Turabian Style

Zhu, Lili, Ying Lin, Wenli Yang, Zhiwen Pan, Weiting Chen, Juan Yao, Ou Sheng, Lingyan Zhou, and Dagang Jiang. 2023. "The Identification of the Banana Endogenous Reference Gene MaSPS1 and the Construction of Qualitative and Quantitative PCR Detection Methods" Genes 14, no. 12: 2116. https://doi.org/10.3390/genes14122116

APA Style

Zhu, L., Lin, Y., Yang, W., Pan, Z., Chen, W., Yao, J., Sheng, O., Zhou, L., & Jiang, D. (2023). The Identification of the Banana Endogenous Reference Gene MaSPS1 and the Construction of Qualitative and Quantitative PCR Detection Methods. Genes, 14(12), 2116. https://doi.org/10.3390/genes14122116

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