DNA isolation from dry samples of broomrape - The effect of isolation method and sample storage on DNA yield and quality

Abstract

Accurate characterization of broomrape races is of a great importance for creation of sunflower hybrids resistant to this parasite. In molecular diversity studies, it is necessary to isolate sufficient quantity of good quality DNA. As collecting and transporting fresh broomrape tissue and seed samples for molecular analyses can be problematic, we have tested the possibility of DNA isolation from dry tissue of mature broomrape plants, with the aim to find optimal method for sample storage and isolation of sufficient quantity of DNA for multiple PCR reactions needed for diversity studies. Mid parts of dry broomrape stalks were collected and stored in four different ways. Ten DNA isolation protocols were tested and the obtained results compared. The best results regarding DNA yield and quality were obtained with dry samples stored at room temperature in ventilated space. DNA suitable for RAPD analysis was isolated with three of ten tested methods. As protocol of Somma (2006) is labour intensive and produced the lowest DNA yield, only DNA isolation with DNeasy® Plant Mini Kit, Qiagen and protocol by Rogers and Bendich (1985) could be recommended for future studies.