A generalized procedure to prepare PCR-grade DNA from different Corylus avellana L. plant tissues for parentage analysis.

Abstract

A generalized protocol has been developed for an easy DNA purification from different hazelnut tissues. The extraction method avoids the use of liquid nitrogen, requires minimal amount (50 mg) of plant tissue, and allows the recovery of 0.8-1.3 μg high molecular weight and PCR-grade DNA from cotyledons and leaves and 0.2-0.3 μg of DNA from the pollen. The purified DNA has the quality to PCR-amplify and is neatly visualized in agarose gels. This protocol could improve and speed-up not only parentage analysis but also the application of marker assisted selection in hazelnut breeding programs, embryo genotyping before seed germination, analysis of the segregation pattern in the seed-progeny originating from the same mother plant, and detection of association among genetic molecular markers and phenotypic traits of cotyledons and embryo axis. Extension of the described procedure to molecular analysis of other difficult tree species is conceivable.