Plant DNA Isolation: A Method to Efficiently Remove Polyphenolics, Polysaccharides, and RNA

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 restric...

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Bibliographic Details
Published in:Taxon 1995-08, Vol.44 (3), p.379-386
Main Authors: Jobes, David V., Hurley, David L., Thien, Leonard B.
Format: Article
Language:English
Subjects:
Centrifugation
Chlorides
DNA
Enzymes
Genomics
Methods and Techniques
Plant DNA
Polymerase chain reaction
Polysaccharides
RNA
Taxa
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Summary: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.
ISSN:0040-0262
1996-8175
DOI:10.2307/1223408