RRC ID 27845
Author Kim BH, Ramanan R, Cho DH, Choi GG, La HJ, Ahn CY, Oh HM, Kim HS.
Title Simple, rapid and cost-effective method for high quality nucleic acids extraction from different strains of Botryococcus braunii.
Journal PLoS ONE
Abstract This study deals with an effective nucleic acids extraction method from various strains of Botryococcus braunii which possesses an extensive extracellular matrix. A method combining freeze/thaw and bead-beating with heterogeneous diameter of silica/zirconia beads was optimized to isolate DNA and RNA from microalgae, especially from B. braunii. Eukaryotic Microalgal Nucleic Acids Extraction (EMNE) method developed in this study showed at least 300 times higher DNA yield in all strains of B. braunii with high integrity and 50 times reduced working volume compared to commercially available DNA extraction kits. High quality RNA was also extracted using this method and more than two times the yield compared to existing methods. Real-time experiments confirmed the quality and quantity of the input DNA and RNA extracted using EMNE method. The method was also applied to other eukaryotic microalgae, such as diatoms, Chlamydomonas sp., Chlorella sp., and Scenedesmus sp. resulting in higher efficiencies. Cost-effectiveness analysis of DNA extraction by various methods revealed that EMNE method was superior to commercial kits and other reported methods by >15%. This method would immensely contribute to area of microalgal genomics.
Volume 7(5)
Pages e37770
Published 2012
DOI 10.1371/journal.pone.0037770
PII PONE-D-11-21428
PMID 22662217
PMC PMC3360624
MeSH Chlorophyta / genetics* Cost-Benefit Analysis DNA, Plant / isolation & purification* Genetic Techniques / economics RNA, Plant / isolation & purification*
IF 2.776
Times Cited 13
WOS Category BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Resource
Algae NIES-836