Chloroplast DNA extraction from herbaceous and woody plants for direct restriction fragment length polymorphism analysis

The technique described here is a fast and simple method of extracting chlo roplast DNA (cpDNA). If overcomes the need for differential centrifugation using density gradients. The leaves do not have to be kept in the dark and lyophilized before extraction, but lyophilization is still possible. The ch loroplasts are specifically lysed in a cell extract of leaves, using a non- ionic detergent. After isolation by centrifugation, the cpDNA is purified b y the combined action of proteolytic enzymes and detergents, followed by th e elimination of proteins using a mixture of chloroform and isoamyl alcohol . This method provided good quality restriction profiles for all species an alyzed.