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基于数字PCR的中黄6106大豆基因组DNA参考物质用于食品和饲料检测的特性分析

Digital PCR-Based characterization of a Zhonghuang 6106 soybean genomic DNA reference material for its food and feed detection.

作者信息

Wang Cheng, Li Ruihuan, Liu Na, Qi Xin, Xu Chao, Yao Junjin, Zhang Chen, Wang Yasi, Lan Qingkuo, Wang Yong, Zhao Xin

机构信息

Tianjin Academy of Agricultural Sciences, Tianjin, 300192, China.

出版信息

Sci Rep. 2025 Oct 3;15(1):34593. doi: 10.1038/s41598-025-18096-6.

Abstract

The genomic DNA reference material of genetically modified soybean Zhonghuang6106, developed in this study, was prepared by extracting genomic DNA from the leaves of both homozygous transgenic soybean Zhonghuang 6106 and non-transgenic soybean Zhonghuang10, followed by proportional mixing of the genomic DNA. Eight qualified laboratories independently validated the certified reference material (CRM) using the digital polymerase chain reaction (dPCR) method. The certified value for the transgene-specific sequence copy number concentration was determined to be (1.04 ± 0.16)×10³ copies·µL, while the copy number ratio of Zhonghuang 6106 to Lectin was 0.047 ± 0.006.Homogeneity and stability assessments revealed that this batch of CRMs exhibited excellent homogeneity and could be stably stored for up to 10 days at 37 °C. Additionally, it remained stable for up to 6 months at -20 °C. This reference material can be used for qualitative and quantitative detection of genetically modified soybean Zhonghuang 6106 and its related products, as well as for the evaluation of specific detection methods and laboratory quality control.

摘要

本研究培育的转基因大豆中黄6106基因组DNA标准物质,是通过提取纯合转基因大豆中黄6106和非转基因大豆中黄10叶片的基因组DNA,然后按比例混合基因组DNA制备而成。八个合格实验室采用数字聚合酶链反应(dPCR)方法对该标准物质进行了独立验证。转基因特异性序列拷贝数浓度的认定值为(1.04 ± 0.16)×10³拷贝·µL,中黄6106与凝集素的拷贝数比为0.047 ± 0.006。均匀性和稳定性评估表明,这批标准物质具有良好的均匀性,在37 °C下可稳定保存长达10天。此外,在-20 °C下可稳定保存长达6个月。该标准物质可用于转基因大豆中黄6106及其相关产品的定性和定量检测,以及特定检测方法的评估和实验室质量控制。

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