Yee Willy, Kumar Janushree Nanda, Muthusamy Priya Devi
Department of Biosciences, Faculty of Applied Sciences, Nilai University, No. 1, Persiaran Universiti, 71800 Nilai, Negeri Sembilan Malaysia.
Indian J Microbiol. 2018 Mar;58(1):109-113. doi: 10.1007/s12088-017-0698-5. Epub 2017 Dec 14.
2-mercaptoethanol (2-ME), alongside polyvinylpyrrolidone is commonly used in plant DNA extractions to deal with polyphenols, which could interfere with extraction and downstream applications. 2-ME is also commonly used to denature proteins and nucleases, especially RNAses. On the contrary, we found that the presence of 2-ME in lysis buffer interfered with DNA extraction from 12 strains of freshwater microalgae, resulting in DNA with poor integrity. We also found that the TNES-urea buffer, commonly used for preservation and DNA extraction from fish, appears as effective as the SDS and CTAB buffer for some microalgae strains. Results from our study suggests that the inclusion of 2-ME in DNA extraction protocols may be detrimental for isolation of good quality DNA from freshwater microalgae, and therefore recommend eliminating it or testing varying concentrations of 2-ME when developing species-specific extraction protocols for microalgae.
2-巯基乙醇（2-ME）与聚乙烯吡咯烷酮一起常用于植物DNA提取中，以处理可能干扰提取及下游应用的多酚类物质。2-ME也常用于使蛋白质和核酸酶变性，尤其是核糖核酸酶。相反，我们发现裂解缓冲液中2-ME的存在会干扰从12株淡水微藻中提取DNA，导致提取的DNA完整性较差。我们还发现，常用于鱼类保存和DNA提取的TNES-尿素缓冲液，对某些微藻菌株而言，似乎与SDS和CTAB缓冲液一样有效。我们的研究结果表明，DNA提取方案中加入2-ME可能不利于从淡水微藻中分离高质量的DNA，因此建议在制定微藻物种特异性提取方案时，去除2-ME或测试不同浓度的2-ME。
