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The use of S1 nuclease treatment of hybrid PCR products for the differentiation between PVY isolates
Author(s) -
ROSNER ARIE,
SPIEGEL SARA,
MASLENIN LUDMILA,
LEVY DAVID
Publication year - 1998
Publication title -
annals of applied biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.677
H-Index - 80
eISSN - 1744-7348
pISSN - 0003-4746
DOI - 10.1111/j.1744-7348.1998.tb05188.x
Subject(s) - biology , nuclease , cleavage (geology) , strain (injury) , potato virus y , polymerase chain reaction , polyacrylamide gel electrophoresis , gel electrophoresis , genetics , microbiology and biotechnology , virology , dna , virus , gene , enzyme , biochemistry , plant virus , paleontology , anatomy , fracture (geology)
Summary A new strategy based on treating PCR hybrids with S1 nuclease was used to differentiate between two PVY isolates. Mixed denatured and annealed hybrid PCR products of two PVY isolates including a tested strain and a reference N strain were treated with S1 nuclease. Single‐stranded mismatched regions were revealed by the S1 nuclease cleavage, yielding a characteristic pattern of bands in polyacrylamide gel by which virus isolates could be matched. Sequence analysis of the relevant PCR products revealed that only part of the mismatched regions were cleaved by the S1 nuclease. Still, the distinct pattern of degradation products enabled the differentiation between the PVY isolates. The general application of this procedure for strain differentiation is discussed.