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Comparative detection of mild strains of potato spindle tuber viroid from the dormant potato tubers by Return-polyacrylamiee gel electrophoresis and nucleic acid hybridization
Authors:R P Singh  A Boucher
Institution:(1) Agriculture Canada, Research Branch, Research Station, P.O. Box 20280, E3B 4Z7 Fredericton, New Brunswick, Canada
Abstract:Summary Using a modified procedure of Return-polyacrylamide gel electrophoresis (R-PAGE) mild strains of potato spindle tuber viroid (M-PSTV) were detected reliably from dormant tubers. The sensitivity of R-PAGE detection of M-PSTV was equivalent to that of nucleic acid hybridization. Both methods detected M-PSTV when infected tissue was mixed with healthy tissue in a ratio of 1 to 100. When extracted nucleic acid was diluted with buffer, R-PAGE detected PSTV at a dilution of 1:256 and nucleic hybridization only up to 1:64 PSTV was readily detected from 18 potato cultivars. In addition, mild, ‘intermediate’ and severe strains were separated by R-PAGE, on the basis of their mobility on the electrophorogram.
Keywords:sensitivity  routine testing  silver staining  dot-blot  severe strain  separation of strains
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