煮沸萃取法与商业试剂盒分离食品样品细菌DNA的比较

M. Dimitrakopoulou, Venia Stavrou, C. Kotsalou, A. Vantarakis
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引用次数: 11

摘要

背景:近年来,商业DNA提取试剂盒的开发,结合分子技术的应用,已被用于了解微生物生态系统。DNA提取被认为是聚合酶链反应等分子技术的关键步骤。目前有几种不同的方法和商业工具包可用于此目的。本研究的目的是比较五种市售试剂盒和一种廉价、快速、非商业的方法——煮沸法,从不同的食物基质中分离细菌DNA。方法:采用实验方案对橄榄、鱼子、葡萄干和培根四种不同类型的食物进行实验。首先用分光光度计测定提取DNA的纯度和浓度,然后用电泳检查DNA的完整性,最后用pcr扩增对结果进行评估。结果:沸煮法高效、简便、廉价,适用于所有食品类型的PCR扩增。结论:煮沸提取食品中细菌DNA的方法有待进一步探索,以验证和建立。
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Boiling Extraction Method VS Commercial Kits for Bacterial DNA Isolation from Food Samples
Background: In recent years, the development of commercial Kits for DNA extraction, combined with the use of molecular technologies have been utilized for the understanding of microbial ecosystems. DNA extraction considered being a critical step for molecular techniques such as Polymerase Chain Reaction. Several different methods and commercial kits for this purpose are nowadays available. The aim of this study was to compare five commercially available kits and, a cheap, rapid, non-commercial method, the boiling method for isolating bacterial DNA from different food matrixes. Methods: Experimental protocol was applied on four different types of food: olives, roe, raisins and bacon. The evaluation of the results was performed first by using a spectrophotometer, by measuring purity and concentration of the extracted DNA, followed by electrophoresis where the integrity of the DNA was checked and finally by PCR-amplification. Results: Results revealed that boiling method is efficient, simple, cheap and suitable for PCR amplification for all these food types. Conclusion: Boiling method for bacterial DNA extraction from food should be further explored in order to be validated and established.
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