Ekstraksi DNA dari Daging Segar untuk Analisis dengan Metode Loop-Mediated Isothermal Amplification (LAMP)

Rosy Hutami, Hanifah Bisyri, Sukarno Sukarno, H. Nuraini, R. Ranasasmita
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引用次数: 3

Abstract

DNA extraction is needed in the analysis using the Loop-mediated isothermal amplification (LAMP) method because this method identifies nucleic acids. Some extraction methods that can be selected including commercial kits extraction method and phenol-chloroform extraction method. The purpose of this study was to obtain the best quality DNA extract between the two extraction methods. The DNA extraction process produced DNA concentrations between 31.06 - 410.18 ng / ml for the commercial kit DNA extract and 212.60 - 1502.30 ng / ml for the phenol-choroform DNA extract, while the purity of DNA were 1.82-2.02 for commercial kit DNA extract and 1.93-2.02 for phenol-chloroform DNA extract. The concentration and purity of extracts produced from both methods meet the requirements for molecular analysis. The purity and visualization results of commercial kit DNA extract are better than those produced from extraction from the phenol-chloroform method. DNA extract obtained from the commercial kit method was chosen to be used in the amplification stage of the method (LAMP).
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使用环介导等温扩增(LAMP)方法进行分析时需要提取DNA,因为该方法鉴定核酸。可选择的提取方法包括商业试剂盒提取法和苯酚-氯仿提取法。本研究的目的是在两种提取方法中获得质量最好的DNA提取物。脱氧核糖核酸(DNA)的萃取浓度为31.06 ~ 410.18 ng / ml,苯酚-氯仿脱氧核糖核酸(DNA)的萃取浓度为212.60 ~ 1502.30 ng / ml,脱氧核糖核酸(DNA)的纯度为1.82 ~ 2.02,苯酚-氯仿脱氧核糖核酸(DNA)的纯度为1.93 ~ 2.02。两种方法提取的提取物浓度和纯度均满足分子分析的要求。商品试剂盒DNA提取液的纯度和可视化结果优于苯酚-氯仿法提取液。选择从商业试剂盒法获得的DNA提取物用于该方法(LAMP)的扩增阶段。
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