An efficient method of DNA extraction from Theobroma cacao and Cola nitida leaves. In: Agrotrópica v. 15 p. 127-130, 2003.

Abstract

An efficient procedure for DNA extraction from recently matured green leaves of Theobroma cacao L. and Cola nitida is described. The procedure has been successfully used for extraction of good quality genomic DNA from cocoa accessions and the related genus, Cola nitida. It is a modified CTAB procedure, which includes use of high concentration of hexadecyltrimethyl ammonium bromide, (CTAB) (4%), and PVP in the extraction buffer to remove polysaccharides and polyphenols respectively. Addition of 5M NaCl during DNA precipitation with isopropanol, use of buffer in higher proportion in the extraction buffer, additional centrifugation steps during chloroform: isoamyl extractions, extended RNAse treatment, additional phenol: chloroform: isoamyl extractions and profuse additional washes of DNA pellet with ethanol to eliminate salt residues further helped in eliminating residual viscous substances and enhancing DNA recovery. The isolated DNA is of good quality and amenable to Polymerase Chain Reaction analysis.