Selection and optimization of the method for DNA isolation and purification from Corylus species for PCR analysis

Abstract

Aim.The use of molecular genetic analysis tools requires DNA preparations of high purity. However, in the case of hazelnut plants (Corylus spp.), obtaining quality DNA is complicated by the presence of a significant amount of secondary metabolites in the leaf tissue. The aim of the study was to select and optimize the method for isolation and purification of hazelnut DNA for further use in PCR analysis. Methods. Three methods of DNA isolation using cetavlon as a detergent were used, which differ in the DNA purification steps: the commonly used protocol by Doyle, Doyle, 1987; a modified method with purification using silicon dioxide microparticles; and a method using the extraction of soluble metabolites, which precedes the DNA isolation step. Results. The first two methods were shown to be unsuitable for DNA isolation from dry hazelnut leaves due to a high amount of impurities. The last method allowed to remove a significant part of water-soluble organic substances at the first stage and obtain high-quality DNA preparations. Conclusions. The method for DNA isolation from Corylus spp. plants and hybrids was selected and optimized, which allows isolation of a sufficient amount of high-quality DNA from dried leaf tissue, suitable for further molecular genetic analysis using PCR.