A simple method for large-scale purification of nucleic acids from plants using calcium phosphate-type monetite

Abstract

Curently, the literature describes several nucleic acids purification methods, depending on the application and the required level of purity. These methods range from simple to complex and are mostly adapted for relatively small scale preparations. As an alternative, we developed in the present work an efficient, rapid, and up-scalable nucleic acids purification method based on the synthesis of a solid Calcium Phosphate-Type Monetite support (CPTM). The synthesis of the CPTM was optimized with regards to the calcium/phosphate (Ca/P) ratio and to sonication parameters (amplitude and time), and phase purity was resolved using X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). Analysis revealed the crystalline purity of the monetite phase and the identity of the matrix, and showing no secondary phases. Nucleic acids adsorption to the CPTM matrix was assessed under optimal conditions of buffering, ionic strength, pH, and flow rate, and the elution was carried out through a phosphate ions gradient that allowed an earlier elution of contaminants. We applied this purification method on several plants material, and results demonstrate that CPTM is a good matrix for nucleic acids purification from complex biological and environmental samples