Abstract
A simplified dot-blot hybridization protocol for Potato spindle tuber viroid (PSTVd) detection in Solanaceae species is described here. The protocol uses an RNA DIG-labeled probe and a simplified extraction procedure that avoids the use of hazardous chemicals. PSTVd was detected in composite tomato leaf samples in a ratio of up to 1:15 of PSTVd-infected to non-infected tissue and in composite potato tuber samples in a ratio up to 1:5 of PSTVd-infected to non-infected tissue. In Brugmansia spp., PSTVd was detected solely in the standard sample extract preparation. The method is suitable for a reliable, large-scale sample screening especially where cost is a limiting factor.
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References
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Vassilakos Ν, Kektsidou O, Papaeythimiou Μ, Varveri C (2012) Comparison of direct-RT-PCR and dot-blot hybridization for the detection of Potato spindle tuber viroid in natural host plant species. Eur J Plant Pathol 134(4):859–864. https://doi.org/10.1007/s10658-012-0061-y
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Kekstidou, O., Varveri, C., Vassilakos, N. (2022). A Simplified Dot-Blot Hybridization Protocol for Potato spindle tuber viroid Detection in Solanaceae. In: Rao, A.L.N., Lavagi-Craddock, I., Vidalakis, G. (eds) Viroids. Methods in Molecular Biology, vol 2316. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1464-8_9
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DOI: https://doi.org/10.1007/978-1-0716-1464-8_9
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1463-1
Online ISBN: 978-1-0716-1464-8
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