Inclusion of 2-Mercaptoethanol in Lysis Buffer Could Interfere with Isolation of High Molecular Weight DNA from Freshwater Microalgae

被引:3
|
作者
Yee, Willy [1 ]
Kumar, Janushree Nanda [1 ]
Muthusamy, Priya Devi [1 ]
机构
[1] Nilai Univ, Dept Biosci, Fac Sci Appl, 1 Persiaran Univ, Nilai 71800, Negeri Sembilan, Malaysia
关键词
2-Mercaptoethanol; DNA extraction; Microalgae; Lysis buffer; TNES-urea; RNA; IDENTIFICATION; EXTRACTION; GROWTH;
D O I
10.1007/s12088-017-0698-5
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:109 / 113
页数:5
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