Rapid Single Nucleotide Polymorphism Genotyping Using FTA Cards and High Resolution Melt Analysis.

Molecular biology tools are often designed to be applicable in the lab to provide answers to specific research questions.  However, many of the techniques cannot be readily applied as high throughput low cost approaches that would have utility for commercial industries or certification agencies.  In genetic marker analysis two areas that increase costs and slow throughput are DNA extraction and fragment analysis.  In an effort to address these problems we have tested the utility of rapid DNA extraction utilizing FTA cards and rapid marker scoring utilizing High Resolution Melt (HRM) analysis in a range of turfgrass genera.

In this study three grass species (Bouteloua dactyloide, Agrostis stolonifera,and Danthonia spicata) were tested. DNA was extracted and stored using FTA cards followed by primer specific PCR and high resolution melt analysis. Concentrations of DNA extracted via FTA card were quantified by a Nano Drop and found to be at satisfactory concentrations for successful amplification using PCR. Differences in melt temperatures as small as 0.25^o C indicated the existence of SNPs. The FTA cards containing transferred plant material were stored in a dry (humidity controlled) area where they can last several months without deterioration. Results show that FTA cards can provide researchers with an effective and efficient tool for collecting storing and extracting grass DNA. The results indicate that FTA card DNA extraction combined with high resolution melt analysis after PCR can be used for rapid SNP genotyping in several turfgrass species.