Servicio de Información Comunitario sobre Investigación y Desarrollo - CORDIS

Protocols for the isolation of SSR markers for Fagaceae and report on their segregation

Primers for the amplification of microsatellites were obtained by sequencing clones of an enriched library constructed for Fagus sylvatica. Microsatellites are markers displaying high variation and extremely useful for several applications in population and conservation genetics, in particular for the analysis of gene flow. Moreover, these markers can be applied for certification purposes, in particular of provenances and/or seed lots and wood material. These new microsatellites developed for beech are already available to scientists and end users thanks to scientific publications where all details about their characteristics and protocols for their screening are reported. Details about the methodology for the amplification of microsatellite markers are provided in a scientific publication. This publication includes: 1) composition of the amplification reaction; 2) thermal profile for the amplification; 3) methodology to visualize SSRs; 4) characteristics of the SSRs. The protocol was developed for the amplification of microsatellites having characteristics necessary to correctly perform paternity analysis, namely absence of null alleles, reproducibility and lack of linkages. Considering that microsatellites useful for paternity analysis must not be linked and must be Mendelian inherited, a segregation and linkage analysis was performed. Mendelian inheritance and absence of linkages between markers was confirmed. Markers having these characteristics have several useful applications in different fields of forest genetics.

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