Chloroplast DNA Extraction from Herbaceous and Woody Plants for Direct Restriction Fragment Length Polymorphism Analysis

Autor: C. Mariac, P. Trouslot, C. Poteaux, G. Bezançon, J.-F. Renno
Jazyk: angličtina
Rok vydání: 2000
Předmět:
Zdroj: BioTechniques, Vol 28, Iss 1, Pp 110-113 (2000)
Druh dokumentu: article
ISSN: 1940-9818
0736-6205
DOI: 10.2144/00281st07
Popis: The technique described here is a fast and simple method of extracting chloroplast DNA (cpDNA). It 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 chloroplasts are specifically lysed in a cell extract of leaves, using a non-ionic detergent. After isolation by centrifugation, the cpDNA is purified by the combined action of proteolytic enzymes and detergents, followed by the elimination of proteins using a mixture of chloroform and isoamyl alcohol. This method provided good quality restriction profiles for all species analyzed.
Databáze: Directory of Open Access Journals