Single-strand conformation polymorphism of hyper-variable regions HV1 and HV2 of human mitochondrial DNA: detection by silver staining.
Human mitochondrial DNA from 50 trios consisting of mother (M), child (C) and father (F) was PCR amplified with primers flanking the hyper-variable regions, HVR1 and HVR2. The amplified products were then fractionated under non-denaturing conditions, silver-stained and compared by single-stranded conformational polymorphism (SSCP). In all but one case, mother and child displayed identical patterns, which could be promptly distinguished from that of the father. For the remaining cases, either set of primers was sufficient to resolve the familial ties. In no instance, M displayed alleles different from those of C within each trio, demonstrating that no false exclusions occurred. The SSCP approach proved to be a robust technique suitable as a preliminary screening in cases requiring identification of multiple samples.
de Souza Menezes
Jackson
J
Departamento de Bioquímica Médica ICB/CCS, Universidade Federal do Rio de Janeiro, Bloco E Sala 22, Ilha do Fundão, Rio de Janeiro, CEP 21941-590, Brazil.
de Almeida Drummond Franklin
Daniela
D
Seki
Homiko
H
Rumjanek
Franklin David
FD
D002648:Child; D016172:DNA Fingerprinting / Q000379:methods; D004272:DNA, Mitochondrial / Q000032:analysis*; D005260:Female; D006801:Humans; D008297:Male; D010334:Paternity; D010375:Pedigree; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D016622:Silver Staining
D004272:DNA, Mitochondrial