Identification of apolipoprotein E polymorphism by using synthetic oligonucleotides.
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.
Smeets
H J
HJ
Department of Human Genetics, Radboud Hospital, University of Nijmegen, The Netherlands.
Poddighe
J
J
Stuyt
P M
PM
Stalenhoef
A F
AF
Ropers
H H
HH
Wieringa
B
B
D001057:Apolipoproteins E / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005784:Gene Amplification; D006801:Humans; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides; D011110:Polymorphism, Genetic; D015183:Restriction Mapping
D001057:Apolipoproteins E; D009841:Oligonucleotides; D004247:DNA