A protocol for high-quality genomic DNA extraction from legumes

• DOI: 10.4238/2012.september.14.1
• Publication Date: 2012-09-17T12:00:18Z
• Authors: I.O. Agbagwa; S. Datta; P.G. Patil; P. Singh; N. Nadarajan
• Keywords: Isoamyl alcohol; Cajanus; genomic DNA; Absorbance

Current DNA extraction protocols, which require liquid nitrogen, lyophilization and considerable infrastructure in terms of instrumentation, often impede the application biotechnological tools less researched crops laboratories developing countries.We modified optimized existing CTAB method for plant genomic by avoiding nitrogen usage lyophilization.DNA was extracted directly from freshly harvested leaves ground pre-heated buffer.Chloroform:isoamyl alcohol (24:1) RNase treatments followed single-purification step decontaminated samples thereby paving way selective DNA.High molecular weight yield range 328 to 4776 ng/µL with an average 1459 obtained 45 cultivated wild Cajanus species.With absorbance ratio at 260 280 nm, a 1.66 2.20, mean 1.85, very low levels protein polysaccharide contamination were recorded.Forty can be daily cost between 1.8 US$2.0 per sample.This is suitable most plants especially members Leguminosae.Apart Cajanus, it has been extensively applied Cicer Vigna species.