基于培养时间的鲜肉(大鼠肉和鸡肉)DNA提取方法优化

Hadi Sunaryo, Adia Putra Wirman, Etin Diah Permanasari, Nurul Azmah Nikmatullah, Dian Lestari, Desi Nurjanah
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引用次数: 0

摘要

DNA(脱氧核糖核酸)提取法是从样品中分离DNA的过程。在这个过程中,必须保护获得的DNA不受RNA、碳水化合物、脂质和蛋白质的污染。RNA、碳水化合物、脂质和蛋白质的污染可以提高DNA的纯度。DNA纯度采用NanoDrop 2000分光光度计测定260 nm和280 nm波长处的吸光度比。优质DNA的A260/A280比值为1.7-2.0,浓度为>本研究旨在为鲜肉样品(大鼠肉和鸡肉的混合物)获得合适的DNA提取方法。本研究包括两个阶段:使用Progenus EasyFast™肉制品提取试剂盒的DNA提取阶段和使用EasyFast™大鼠检测试剂盒的扩增阶段。本研究使用了16份鼠肉和鸡肉的混合物样品,鼠肉的浓度分别为:5%、10%、15%和20%。在提取阶段,优化孵育时间为15、30、45分钟和1小时。结果表明,1 h的PCR扩增结果呈s型曲线。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Optimization of DNA Extraction Methods in Fresh Meat (Rat and Chicken Meat) based on Incubation Time
DNA (deoxyribonucleic acid) extraction method is the process of separating DNA from the sample. In this process, the DNA obtained must be protected from contamination by RNA, carbohydrates, lipids, and proteins. Contamination of RNA, carbohydrates, lipids, and proteins can increase DNA purity. DNA purity was measured using a NanoDrop 2000 spectrophotometer measured by the absorbance ratio at 260 nm and 280 nm wavelengths. Good quality DNA will have an A260/A280 ratio of 1.7–2.0 and a concentration > 0.03 pg. This study aimed to obtain the appropriate DNA extraction method for fresh meat samples (a mixture of rat and chicken meat). This research consisted of two stages: the DNA extraction stage using the Progenus EasyFast™ Extraction Kit for Meat Products and the amplification stage using the EASYFAST™ Rat Detection Kit. This study used 16 samples of a mixture of rat meat and chicken with concentrations of rat meat: 5, 10, 15, and 20%. At the extraction stage, the incubation time was optimized for 15, 30, 45 minutes, and 1 hour. The results showed that the one hour incubation had a sigmoid curve in the results of PCR amplification.
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