Abstract
The high polysaccharide content of some plant species hinders the successful isolation of their DNA. As an alternative to the macro-extraction methods previously published for polysaccharide-rich plants, we present two techniques (STE/CTAB and HEPES/CTAB), which are performed in microcentrifuge tubes. These protocols are suitable for small amounts of silica gel-preserved plant tissue such as are commonly available from endangered plants. The critical step to remove polysaccharides was performing initial washes in either STE (0.25 M sucrose, 0.03 M Tris, 0.05 M EDTA) or HEPES (2% β-mercaptoethanol, 0.2% PVP, 0.1 M HEPES, pH 8.0) buffer. Precipitating the DNA at room temperature with isopropanol also aided in decreasing polysaccharide co-precipitation. Of the two protocols we present the STE/CTAB method has the advantages of being more cost-effective and avoiding the use of the hazardous chemical β-mercaptoethanol.
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Acknowledgments
This study was funded by the New Zealand Marsden Fund (contract number MAU0709). We thank Peter Lockhart and Leon Perrie for comments on the manuscript.
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Shepherd, L.D., McLay, T.G.B. Two micro-scale protocols for the isolation of DNA from polysaccharide-rich plant tissue. J Plant Res 124, 311–314 (2011). https://doi.org/10.1007/s10265-010-0379-5
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DOI: https://doi.org/10.1007/s10265-010-0379-5