DNA Extraction from Moth bean mutants containing high polyphenols and polysaccharides
Keywords:
Moth bean, mutants, DNA extraction, PCR and RAPDAbstract
A number of methods are available for isolation of plant genomic DNA, but in these procedures due to variability in plant tissue composition (like polyphenols, tannins and polysaccharides) practical difficulties are always met. With consistent isolation of quality DNA from Moth bean (Vigna aconitifolia (Jacq.) Marachel) is particularly problematical due to the presence of phenolic compounds. Inconsis-tencies in extraction result can be attributed to the growth stages of the plant material analyzed. The mature moth bean leaves which contain high levels of polyphenols, tannins and polysaccharides, were used as a study group. The method involves a modified CTAB extraction employing high salt concentration to remove polysaccha-rides, the use of polyvinyl pyrrolidone (PVP) to remove polyphenols, an RNase treatment and a phenol-chloroform and chloroform-isoamyl alcohol extraction procedure. The results from the local Moth bean-88 and its seven mutants were consistently amplifiable in RAPD reaction with as little as l µg DNA/25ul reaction.
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