Literature DB >> 29434405

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

Willy Yee1, Janushree Nanda Kumar1, Priya Devi Muthusamy1.   

Abstract

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.

Entities:  

Keywords:  2-Mercaptoethanol; DNA extraction; Lysis buffer; Microalgae; TNES-urea

Year:  2017        PMID: 29434405      PMCID: PMC5801187          DOI: 10.1007/s12088-017-0698-5

Source DB:  PubMed          Journal:  Indian J Microbiol        ISSN: 0046-8991            Impact factor:   2.461


  12 in total

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2.  Feasibility of various carbon sources and plant materials in enhancing the growth and biomass productivity of the freshwater microalgae Monoraphidium griffithii NS16.

Authors:  Willy Yee
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Journal:  Am J Bot       Date:  2007-05       Impact factor: 3.844

5.  LSU rDNA based RFLP assays for the routine identification of Gambierdiscus species.

Authors:  Yihua Lyu; Mindy L Richlen; Taylor R Sehein; Mireille Chinain; Masao Adachi; Tomohiro Nishimura; Yixiao Xu; Michael L Parsons; Tyler B Smith; Tianling Zheng; Donald M Anderson
Journal:  Harmful Algae       Date:  2017-05-18       Impact factor: 4.273

6.  Evaluation of DNA extraction methods for freshwater eukaryotic microalgae.

Authors:  Lucy E Eland; Russell Davenport; Cesar R Mota
Journal:  Water Res       Date:  2012-07-24       Impact factor: 11.236

7.  Effects of reducing and oxidizing agents on the action of bleomycin.

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Journal:  J Biochem       Date:  1975-04       Impact factor: 3.387

8.  Isolation of RNA of high quality and yield from Ginkgo biloba leaves.

Authors:  Tao Wang; Nianhui Zhang; Lingfang Du
Journal:  Biotechnol Lett       Date:  2005-05       Impact factor: 2.461

9.  An efficient protocol for the Agrobacterium-mediated genetic transformation of microalga Chlamydomonas reinhardtii.

Authors:  P T Pratheesh; M Vineetha; G Muraleedhara Kurup
Journal:  Mol Biotechnol       Date:  2014-06       Impact factor: 2.695

10.  CHANGES IN ENDOGENOUS CYTOKININ CONCENTRATIONS IN CHLORELLA (CHLOROPHYCEAE) IN RELATION TO LIGHT AND THE CELL CYCLE(1).

Authors:  Wendy A Stirk; Johannes van Staden; Ondřej Novák; Karel Doležal; Miroslav Strnad; Petre I Dobrev; György Sipos; Vince Ördög; Péter Bálint
Journal:  J Phycol       Date:  2011-03-01       Impact factor: 2.923

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