不同DNA提取方法的比较评价及市售栗子玫瑰果汁和饮料中DNA降解水平的分析。

IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY BMC Biotechnology Pub Date : 2025-01-21 DOI:10.1186/s12896-024-00933-7
Yongchao Ren, Yunlong Ma, Yanqi Li, Yun Song, WeiWei Zhao, Xuncai Huang, Danmin Yu, Jian Li, Zuogang Xu, Wenjun Zhao
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引用次数: 0

摘要

背景:食品安全是一个重要的全球性研究课题,与人类的生活和福祉密切相关。利用基于dna的方法进行物种鉴定在食品检验和监管领域是一种有价值的工具。它对于可追溯性的目的尤其重要,因为它可以在食品链监管的每个级别对特定项目进行监控。然而,在此过程中获得可扩增的基因组DNA是基因研究的一个重大障碍。迄今为止,缺乏从加工果汁或饮料中提取DNA的文献,也没有同时比较各种提取工艺的数据。本研究旨在优化和比较栗子玫瑰果汁和饮料中4种DNA提取方法。此外,我们还利用扩增子大小对板栗玫瑰汁或饮料中的DNA降解程度进行了比较和分析。方法:采用NanoDrop One分光光度计、凝胶电泳、实时聚合酶链反应(real-time PCR或qPCR)等方法评价提取DNA的数量和质量。对每种方法的处理时间、劳动强度和成本进行了评估。采用TaqMan实时PCR方法对板栗玫瑰汁或饮料中的DNA降解程度进行了评估。结果:非商业改良的基于ctab的方法产生了高的DNA浓度。然而,分光光度和实时PCR分析结果显示DNA质量较差。该方法在提取方法中表现出最好的性能,但与其他方法相比,耗时和成本较高。此外,DNA降解的分析结果表明,样品DNA的完整性可能受到不同制造商使用的复杂处理方法的影响。结论:为了实现精确的DNA定量,有必要针对给定的基质选择合适的提取策略。该组合法是最有效的DNA提取技术,可用于板栗玫瑰果汁和饮料中DNA的提取。这种比较评估对于提取和识别各种食品成分中的加工果汁和饮料特别有价值。
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Comparative evaluation of various DNA extraction methods and analysis of DNA degradation levels in commercially marketed Chestnut rose juices and beverages.

Background: Food safety is a significant global study subject that is strongly intertwined with human life and well-being. The utilization of DNA-based methods for species identification is a valuable instrument in the field of food inspection and regulation. It is particularly significant for traceability purposes, as it enables the monitoring of a specific item at every level of the food chain regulation. However, obtaining amplifiable genomic DNA in this process is a significant obstacle in gene studies. To date, there is a lack of literature on DNA extraction from processed juice or beverages, and no data exist on simultaneous comparisons of various extraction processes. This study aimed to optimize and compare four DNA extraction methods for Chestnut rose juices and beverages. Furthermore, we also conducted a comparison and analysis of the extent of DNA degradation in Chestnut rose juice or beverage by utilizing the amplicon size.

Methods: The quantity and quality of the extracted DNA were assessed using NanoDrop One spectrophotometer, gel electrophoresis, and real-time polymerase chain reaction (real-time PCR or qPCR) assays. An assessment was conducted on the processing time, labor intensity, and cost associated with each approach. The degree of DNA degradation in Chestnut rose juice or beverage was also assessed using TaqMan real-time PCR methods.

Results: The non-commercial modified CTAB-based approach yielded a high DNA concentration. However, spectrophotometric results and real-time PCR analysis showed poor DNA quality. The combination approach showed the greatest performance among the extraction methods, while being comparatively time-consuming and costly in contrast to the other methods. Additionally, the analytical findings of DNA degradation suggested that the integrity of sample DNA could be influenced by the intricacy of processing methods used by various manufacturers.

Conclusions: To achieve precise DNA quantification, selecting suitable extraction strategies for the given matrix is necessary. The combination approach was identified as the most effective DNA extraction technique and is suggested for extracting DNA from Chestnut rose juices and beverages. This comparative assessment can be particularly valuable for extracting and identifying processed Juices and Beverages in a diverse range of food compositions.

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来源期刊
BMC Biotechnology
BMC Biotechnology 工程技术-生物工程与应用微生物
CiteScore
6.60
自引率
0.00%
发文量
34
审稿时长
2 months
期刊介绍: BMC Biotechnology is an open access, peer-reviewed journal that considers articles on the manipulation of biological macromolecules or organisms for use in experimental procedures, cellular and tissue engineering or in the pharmaceutical, agricultural biotechnology and allied industries.
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