An alternative and effective tissue pre-treatment to improve the DNA identification of canned tuna samples

IF 4.6 2区 农林科学 Q2 CHEMISTRY, APPLIED
Maria Clara G. Queiroz-Brito , Paulo Travassos , Mônica L. Adam , Rodrigo A. Torres
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Abstract

DNA purity represents a major challenge to investigate food authenticity of canned products due to DNA degradation. Herein, we present a low-cost protocol to increase concentration and purity of DNA extracted from canned samples. The experiment mainly consists of: (1) drying the canned tissue in paper filter, (2) washing it with a PBS solution, (3) store in ethanol 96 % at −20°C, and (4) perform DNA extraction. The pre-treated samples showed an increase of both DNA concentration and purity (A260/A280 ratio), indicating that some of the inhibiting molecules were successfully removed. These differences between the two treatments were statistically significant (p < 0.01). At the amplification level, the pre-treatment allowed the recovery of complete fragments of the barcode region COX1 with approximately 650 bp. Since obtaining relevant levels of DNA purity and concentration from degraded samples, and the sequencing of large fragments from processed samples represents a difficult task, the presented results demonstrate a positive effect of the proposed protocol. Thus, the combination of this treatment with other methodologies, such as mini-barcoding, and sample types is strongly encouraged.
提高金枪鱼罐头样品DNA鉴定的一种有效的组织预处理方法
由于DNA的降解,DNA纯度是调查罐头食品真实性的主要挑战。在此,我们提出了一种低成本的方案,以提高从罐装样品中提取DNA的浓度和纯度。实验主要包括:(1)用纸滤纸干燥,(2)PBS溶液洗涤,(3)96 %乙醇,- 20℃保存,(4)DNA提取。预处理后的样品显示DNA浓度和纯度(A260/A280比)均有所增加,表明部分抑制分子被成功去除。两组间差异均有统计学意义(p <; 0.01)。在扩增水平,预处理允许恢复条形码区域COX1的完整片段,大约650 bp。由于从降解样品中获得相关水平的DNA纯度和浓度,并且从处理过的样品中对大片段进行测序是一项艰巨的任务,因此所提出的结果表明了所提出方案的积极效果。因此,强烈建议将这种处理方法与其他方法(如迷你条形码和样本类型)相结合。
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来源期刊
Journal of Food Composition and Analysis
Journal of Food Composition and Analysis 工程技术-食品科技
CiteScore
6.20
自引率
11.60%
发文量
601
审稿时长
53 days
期刊介绍: The Journal of Food Composition and Analysis publishes manuscripts on scientific aspects of data on the chemical composition of human foods, with particular emphasis on actual data on composition of foods; analytical methods; studies on the manipulation, storage, distribution and use of food composition data; and studies on the statistics, use and distribution of such data and data systems. The Journal''s basis is nutrient composition, with increasing emphasis on bioactive non-nutrient and anti-nutrient components. Papers must provide sufficient description of the food samples, analytical methods, quality control procedures and statistical treatments of the data to permit the end users of the food composition data to evaluate the appropriateness of such data in their projects. The Journal does not publish papers on: microbiological compounds; sensory quality; aromatics/volatiles in food and wine; essential oils; organoleptic characteristics of food; physical properties; or clinical papers and pharmacology-related papers.
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