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Plant DNA isolation: a method to efficiently remove polyphenolics, polysaccharides, and RNA
Author(s) -
Jobes David V.,
Hurley David L.,
Thien Leonard B.
Publication year - 1995
Publication title -
taxon
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 81
eISSN - 1996-8175
pISSN - 0040-0262
DOI - 10.2307/1223408
Subject(s) - dna , polyphenol , polysaccharide , rna , chemistry , polyvinylpyrrolidone , restriction enzyme , biochemistry , ethidium bromide , polymerase chain reaction , chromatography , biology , gene , organic chemistry , antioxidant
Summary Jobes, D. V., Hurley, D. L. & Thien, L. B.: Plant DNA isolation: a method to efficiently remove polyphenolics, polysaccharides, and RNA. – Taxon 44: 379‐386. 1995. – ISSN 0040‐0262. The isolation of DNA from plants has been described by numerous authors, each contributing a different method to overcome the problems that arise when extracting DNA. There are at least three main contaminants associated with plant DNA that can cause considerable difficulties when conducting restriction enzyme analyses and PCR (polymerase chain reaction) experiments: polyphenolic compounds, polysaccharides, and RNA. The procedure described below alleviates the time and expense of CTAB (cetyltrimethylammonium bromide) and caesium chloride methods by utilizing PVP (polyvinylpyrrolidone) to bind the polyphenolic compounds, a high molar concentration of sodium chloride to inhibit co‐precipitation of the polysaccharides and DNA, and an improved method for removing RNA by selective precipitation with lithium chloride. Isolated DNA was easily digested with restriction enzymes and amplified by the PCR from widely different plant species.