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Georgia Journal of Science

Article Title

CROSS SPECIES TRANSFERENCE OF MICROSATELLITE PRIMERS FOR GENETIC MARKER INDENTIFCATION IN SPANISH MOSS TILLANDSIA USNEOIDES**

Abstract

Tillandsia usneoides (Spanish moss) is an ecologically and culturally important epiphyte. Nevertheless, many questions about Spanish moss dispersal, including the relative frequency of different reproductive modes (i.e., seed dispersal vs. vegetative breakage), have not been resolved. This lack of understanding is in part due to a lack of defined hypervariable DNA-based markers for this species. This may be partly due to the fact that it is difficult to obtain substantial amounts of high-quality DNA. Our study examined cross-species transference of microsatellite primers developed for various species in the family Bromeliaceae to T. usneoides. This was done by screening 36 published microsatellite primer sets by way of agarose gel electrophoresis and using an Agilent 2200 TapeStation. PCR product analyzed using an Applied Biosystems 3730 Genetic Analyzer with Capillary Electrophoresis produced varying results. Primer sets were either completely monomorphic, showed limited variability, or produced electropherograms that were difficult to interpret. In order to confirm these results, DNA extraction protocols were optimized to obtained sufficient amounts of pure DNA which would eliminate potentially inhibitory contaminants that could affect downstream processes (notably, tannins, polysaccharides and residual phenol). Single strands of T. usneoides were extracted to ensure that multiple genetic profiles are not present as suggested by unclear peak patterns. PCR protocols will be optimized in order to ensure correct amplification of the regions of interest and minimize non-specific product amplifications.

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