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Molecular taxonomy using single‐strand conformation polymorphism (SSCP) analysis of mitochondrial ribosomal DNA genes
Author(s) -
Hiss R. H.,
Norris D. E.,
Dietrich C. H.,
Whitcomb R. F.,
West D. F.,
Bosio C. F.,
Kambhampati S.,
Piesman J.,
Antolin M. F.,
IV W. C. Black
Publication year - 1994
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1111/j.1365-2583.1994.tb00164.x
Subject(s) - biology , single strand conformation polymorphism , mitochondrial dna , genetics , gene , polymerase chain reaction , ribosomal rna , ribosomal dna , dna , microbiology and biotechnology , phylogenetic tree
Single‐strand conformation polymorphism (SSCP) analysis detects single point mutations in DNA molecules. We demonstrate that SSCP analysis of mitochondrial ribosomal DNA (rDNA) genes is a sensitive taxonomic tool because these genes often differ at numerous sites among closely related species. Using conserved primers, portions of the 12S or 16S rDNA genes were amplified using the polymerase chain reaction (PCR) in congeneric species of ticks, leaf hoppers, mosquitoes, and closely related endoparasitic wasps. SSCP was performed and products were visualized with silver staining. Species‐specific patterns were observed in all taxa. Intraspecific variation at the level of single nucleotide substitutions was detected. SSCP diagnostics are less expensive and time consuming to develop than PCR with species‐specific primers, and, unlike PCR with arbitrary primers, there is minimal concern with DNA contamination from non‐target organisms.