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Effect of DNA extraction methods on the detection of porcine ingredients in halal cosmetics using real-time PCR

机译:DNA提取方法对使用实时PCR检测清真化妆品猪成分的影响

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In recent years, halal cosmetics have attracted considerable attention worldwide. We developed a real-time PCR assay based on the mitochondrial gene ndh5 for rapid detection of porcine ingredients in halal cosmetic products. We also compared several DNA extraction methods for the most efficient approach in different types of cosmetics. Porcine template DNA was spiked into three types of cosmetics (liquid-type and powder-type mask packs, and cream) and extracted with five commercial DNA extraction kits and the CTAB method. The extraction efficiency of each method was evaluated by determining the detection limits of real-time PCR assay. The lowest detection limit of real-time PCR for each cosmetic product was as follows: 2.28 x 10(0) copies for liquid-type mask pack when the Power Prep (TM) DNA extraction kit and TIANamp Genomic DNA kit were used, 2.28 x 10(1) copies for powder-type mask pack when QIAamp DNA stool mini kit and the Power Prep (TM) DNA extraction kit were used, and 2.28 x 10(0) copies for cream when the Power Prep (TM) DNA extraction kit was used. The pig-specific real-time PCR assay facilitated the detection of trace amounts of the template DNA in cosmetics, and an appropriate DNA extraction method was used depending on the type of cosmetics.
机译:近年来,清真化妆品在全球上引起了相当大的关注。我们基于线粒体基因NDH5开发了实时PCR测定,用于快速检测清真化妆品中的猪成分。我们还将几种DNA提取方法与不同类型化妆品中最有效的方法进行了比较了几种DNA提取方法。猪模板DNA被掺入三种类型的化妆品(液体型和粉末型掩模包装和乳霜)中,并用五种商业DNA提取套件和CTAB方法提取。通过确定实时PCR测定的检测限评估每种方法的提取效率。每个化妆品的实时PCR的最低检测限如下:2.28 x 10(0)液体型掩模包装拷贝,当使用POWER PREP(TM)DNA提取试剂盒和天然基因组DNA试剂盒,2.28 x 10(1)粉末型掩模包装的拷贝QIAAMP DNA粪便迷你试剂盒和功率准备(TM)DNA提取试剂盒,功率准备(TM)DNA提取试剂盒时为2.28×10(0)份。用过。猪特异性实时PCR测定促进了在化妆品中的痕量模板DNA的检测,并且根据化妆品的类型使用了适当的DNA提取方法。

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