PCR Amplification of RNA of IYSV and TSWV recovered and stored on FTA cards
Gitaitis R1, Martinez N2, Seebold K3, Stevenson K1, Sanders H1, Mullis S1.
1Univ. of Georgia, Tifton, GA; 2Lexington, KY, 3Univ. of KY, Lexington, KY USA.
Correspondence: dronion@uga.edu
Tissues from onion leaves infected with Tomato Spotted Wilt Virus (TSWV), or Iris Yellow Spot Virus (IYSV) and from tobacco infected with TSWV were triturated in phosphate-buffered saline (PBS) and 100 ul of the homogenate were spotted onto Whatman FTA cards and allowed to dry. Discs (2¨C3 mm diam) were punched from the FTA card and the bound nucleic acids on the paper discs served as a template for RT-PCR. Amplification products were detected by gel electrophoresis through a 1.0% agarose gel containing ethidium bromide in a 0.04M Tris¨Cacetate 0.001M EDTA (TAE) buffer. Basically constructed of filter paper impregnated with proprietary chemicals that are activated by biological fluids, FTA cards can be used for a variety of purposes ranging from the rapid extraction of nucleic acids from ephemeral samples immediately under field conditions to archiving DNA and RNA samples and storing them at room temperature. Furthermore, since viruses are inactivated, FTA cards can be used for transportation of genetic material without the need for special biohazard precautions or plant pest permits.
