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A method for isolation and purification of peanut genomic DNA suitable for analytical applications
Authors:Kiran K Sharma  M Lavanya  V Anjaiah
Institution:(1) Genetic Transformation Laboratory, Genetic Resources and Enhancement Program, International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), 502 324 Patancheru, Andhra Pradesh, India
Abstract:Numerous methods are available for isolation of plant genomic DNA, but in practice these procedures are empirical due to variability in plant tissue composition. Consistent isolation of quality DNA from peanut (Arachis hypogaea L.) is particularly problematic due to the presence of phenolic compounds and polysaccharides. Inconsistencies in extraction results can be attributed to the age and growth stage of the plant material analyzed. Mature leaves have higher quantities of polyphenols, tannins, and polysaccharides that can contaminate DNA during isolation. We show that four published protocols could not be used to isolate peanut DNA of sufficient quality for PCR amplification or Southern hybridization. We have devised a new protocol that uses DEAE-cellulose purification to isolate peanut DNA useful for downstream applications.
Keywords:DNA isolation  PCR amplification  peanut  Southern hybridization  transgenic plants
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