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Identification of apolipoprotein E polymorphism by using synthetic oligonucleotides
H J Smeets1, J Poddighe, P M Stuyt
1Department of Human Genetics, Radboud Hospital, University of Nijmegen, The Netherlands.
Abstract:
A procedure based on selective hybridization with allele-specific oligonucleotides was developed for typing apolipoprotein E variants from human genomic DNAs. Two sets of oligonucleotides were synthesized and used to discriminate either between epsilon 3 and epsilon 4 alleles or between epsilon 3 and epsilon 2 alleles. Combination of the allele-specific oligonucleotide hybridization with the method for in vitro DNA amplification (Polymerase Chain Reaction) Saiki, R. K. et al. 1985. Science, 230: 1350-1354) (1) dramatically improved the sensitivity and the reliability of the procedure. Adaptation of a simple strategy involving direct cloning and DNA sequencing of in vitro amplified DNA enables rapid identification of any mutation within the apoE gene area encoding the receptor binding domain.