Isolation of High Quality Dna for Aflp Amplification in Casuarina Equisetifolia L

High quality DNA was isolated from Casuarina equisetifolia which is widely planted in tropics and subtropics for its multiple utilities. Two commonly used methods, CTAB based and the commercially supplied DNeasyTM Plant Mini Kit (QIAGEN GmbH, Germany) were tested for their efficiency in isolating high quantity and quality DNA to be utilized for AFLP analysis. The CTAB based protocol was modified by introducing additional steps and manufacturer’s protocol was followed for the commercial kit. DNA yield and purity were monitored by gel electrophoresis and by determining absorbance under UV (A260/A280 and A260/A230). Both ratios were between 1.9 and 2.1, indicating the absence of contaminating metabolites. The average DNA yield obtained from 100 mg needle tissue using the CTAB based protocol and the commercial kit was 17 mg and 6 mg, respectively. Further, the purity of DNA was assessed by enzymatic digestion with EcoRI and HindIII and the DNA was also tested for suitability by AFLP-PCR analysis. A unique AFLP pattern was achieved for the five ortets tested showing the utility of DNA for fingerprint studies. Based on the quantity of isolated DNA, the CTAB based method is recommended for C. equisetifolia DNA extraction and it can be utilized for other Casuarina species with high content of fiber and phenolic compounds.