High throughput fluorescent CE-SSCP SNP genotyping.
Large numbers of single nucleotide polymorphisms (SNPs) are being identified by several laboratories for the purpose of developing dense genetic maps. Single-strand conformation polymorphism (SSCP) analysis has been widely used as a method for detecting novel sequence variations in PCR products. Differences in migration of single-stranded DNA can be used not only to find mutations, but to genotype SNPs in large sample populations. Using PCR with fluorescent labeling and automated capillary electrophoresis SSCP (CE-SSCP), we have developed a panel of 15 functional candidate SNPs. With an automated single capillary instrument, relatively rapid and low cost CE-SSCP SNP genotyping using currently available technology is feasible for 135 000 genotypes per year. With parallel multiple array capillary electrophoresis, more genotypes per year may be attainable.
Gonen
D
D
Laboratory for Developmental Neuroscience, Child and Adolescent Psychiatry, Department of Psychiatry, University of Chicago, MC3077, 5841 S Maryland Avenue, Chicago, Illinois 60637, USA. ed@yoda.bsd.uchi cago.edu
Veenstra-VanderWeele
J
J
Yang
Z
Z
Leventhal
B
B
Cook
E H
EH
D001331:Automation / Q000379:methods; D002874:Chromosome Mapping / Q000379:methods*; D017931:DNA Primers; D019075:Electrophoresis, Capillary / Q000379:methods; D005456:Fluorescent Dyes; D005838:Genotype; D006801:Humans; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic; D018807:Polymorphism, Single-Stranded Conformational; D011506:Proteins / Q000235:genetics
D017931:DNA Primers; D005456:Fluorescent Dyes; D011506:Proteins