Umesha S, Manukumar H M, Raghava Sri
Department of Studies in Biotechnology, University of Mysore, Manasagangotri, Mysore, 570006, Karnataka, India.
3 Biotech. 2016 Dec;6(2):123. doi: 10.1007/s13205-016-0436-4. Epub 2016 Jun 6.
Food contaminated with fungal pathogens can cause extremely harmful effects to human even when present in low concentrations. Researchers now pay more attention towards rapid DNA extraction for the quick screening, which is highly demanded in diverse research field. Molecular description of many fungal species is identified by different molecular characteristics. Hence, the efficient isolation of genomic DNA and amplification using PCR is a prerequisite for molecular characterization. Here, we used an improved Sodium dodecyl sulfate-Cetyltrimethyl ammonium bromide-Chloroform-isoamyl alcohol method by combining Sodium dodecyl sulfate with cetyl methylammonium bromide without addition of proteinase K, RNase K, and β-mercaptoethanol. To analyze the quality of recovered DNA, this method was compared with the other four routine methods. The present method has been chosen in the study as a preferred method because of easy adaptation to routine laboratories/food industries considering its rapid, sensitivit,y and cost effectiveness involved in the method.
受真菌病原体污染的食物即使含量很低也会对人类造成极其有害的影响。研究人员现在更加关注用于快速筛查的快速DNA提取，这在不同研究领域都有很高的需求。许多真菌物种的分子描述是通过不同的分子特征来确定的。因此，高效分离基因组DNA并使用PCR进行扩增是分子表征的前提条件。在这里，我们使用了一种改进的十二烷基硫酸钠-十六烷基三甲基溴化铵-氯仿-异戊醇方法，将十二烷基硫酸钠与十六烷基甲基溴化铵结合，不添加蛋白酶K、核糖核酸酶K和β-巯基乙醇。为了分析回收DNA的质量，将该方法与其他四种常规方法进行了比较。由于该方法具有快速、灵敏且成本效益高的特点，易于应用于常规实验室/食品行业，因此在本研究中被选为首选方法。
