Purification of DNA from chloroplast and mitochondria of sugarcane

Abstract

We report here a method for the isolation of chloroplast DNA (ctDNA) and mitochondrial DNA (mtDNA) from leaf samples of 12 elite Indian sugarcane cultivars. This method includes isolation of organelles, purification by deoxyribonuclease treatment, organelle lysis, isolation of nucleic acid with phenol-chloroform and a final DNA purification using CTAB treatment. The protocol offers pure and completely restrictable ctDNA and mtDNA without nuclear DNA contamination. Inter-organellar DNA contamination was confirmed by PCR amplification using respective organellar DNA-specific universal primers. The ctDNA and mtDNA yield was approximately 7 and 5 ?g/g of leaf sample respectively. The method is useful for the isolation of ctDNA and mtDNA from plants that secrete high levels of polysaccharides and phenolic compounds.