title: SNaPshot and CE-SSCP: Two Simple and Cost-Effective Methods to Reveal Genetic Variability Within a Virus Species.
abstract: The multiplex SNaPshot and the capillary electrophoresis-single-strand conformation polymorphism (CE-SSCP) procedures are here used for rapid and high-throughput description of the molecular variability of viral populations. Both approaches are based on (1) standard amplification of genomic sequence(s), (2) labeled primers or labeled single-stranded DNA, and (3) migration of fluorescent-labeled molecules in capillary electrophoresis system. The SNaPshot technology was used to describe the diversity of 20 targeted single nucleotide polymorphisms (SNPs) selected from alignment of viral genomic sequences retrieved from public database. The CE-SSCP procedure was applied to identify the polymorphisms of two small (<500 bases in length) genomic regions of viral genomes. The different steps of SNaPshot and CE-SSCP setup procedures are presented using Potato virus Y (PVY, Potyvirus) and Plum pox virus (PPV, Potyvirus) RNA viruses as molecular targets, respectively.
authors.affiliation: INRA-Cirad-Montpellier SupAgro, UMR 385 BGPI, Cirad TA A-54K, Montpellier cedex, 34398, France.
authors.forename: Agnès
authors.initials: A
authors.lastname: Delaunay
authors.forename: Sylvie
authors.initials: S
authors.lastname: Dallot
authors.forename: Denis
authors.initials: D
authors.lastname: Filloux
authors.forename: Virginie
authors.initials: V
authors.lastname: Dupuy
authors.forename: Philippe
authors.initials: P
authors.lastname: Roumagnac
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Jacquot
mesh_terms: D003362:Cost-Benefit Analysis; D017931:DNA Primers / Q000737:chemistry; D004277:DNA, Single-Stranded / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D019075:Electrophoresis, Capillary / Q000191:economics / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D018807:Polymorphism, Single-Stranded Conformational / Q000235:genetics*; D017800:Potyvirus / Q000145:classification* / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000191:economics / Q000379:methods*
chemical_list: D017931:DNA Primers; D004277:DNA, Single-Stranded; D004279:DNA, Viral