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Microalgae Direct Extract Reagent for Heterocapsa triquetra

  • Gna Ahn
  • Ga-Young Park
  • Dae-Young Park
  • Ok Chan Jeong
  • Yang-Hoon KimEmail author
  • Ji-Young AhnEmail author
Article
  • 3 Downloads

Abstract

Objective

Heterocapsa triquetra (H. triquetra) is classified as bloom-forming dinoflagellate. Early detection is needed to prevent damage by them in the field. Cells of H. triquetra are more difficult to break than bacteria or animal cells because their cells are covered by amorphous cellulose. Therefore, it is necessary to manufacture algae specific buffer that can easily and quickly lyse H. triquetra.

Methods

In the present study, we made lysis buffer to extract DNA and protein from H. triquetra with treatment at room temperature for several minutes. As a result, H. triquetra cells were lysed and dual quantification of DNA and protein could be performed. To confirm DNA extraction, internal transcribed spacer (ITS) region PCR was performed. For protein extraction, SDS-PAGE followed by silver staining and ATPase activity test were used.

Results

Results showed that lysis buffer containing 1.0–5.0% of SDS was efficient for gDNA and protein extraction. The presence of 1 mM EDTA in the buffer was key factor affecting cell wall loosening during H. triquetra cell lysis.

Conclusion

Our MDE method is rapid and simple. It is useful for other microalgae studies.

Keywords

Heterocapsa triquetra Algae Lysis buffer 

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Notes

Acknowledgements

This work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (PJ011661)” funded by Rural Development administration, Republic of Korea. This research was also supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (NRF-2017R1A2B4012253).

Supplementary material

13530_2019_390_MOESM1_ESM.pdf (197 kb)
Microalgae Direct Extract Reagent for Heterocapsa triquetra

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Copyright information

© The Korean Society of Environmental Risk Assessment and Health Science and Springer 2019

Authors and Affiliations

  1. 1.School of Biological SciencesChungbuk National UniversityCheongjuRepublic of Korea
  2. 2.HPBio LaboratorySeoulRepublic of Korea
  3. 3.Department of Biomedical EngineeringInje UniversityGimhaeRepublic of Korea

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