Genetic analysis of the polymorphism of the human apolipoprotein E using automated solid-phase sequencing.
A direct sequencing approach has been used to analyze the polymorphism in the human apolipoprotein E gene. A method is described, in which the DNA is amplified by the polymerase chain reaction, immobilized, and sequenced by a semi-automatic procedure adaptable to clinical diagnosis. The three alleles of the apolipoprotein E gene, which differ from each other by two nucleotide substitutions and which influence serum cholesterol levels, were analyzed. The solid-phase method was able to resolve the correct nucleotide sequence in samples from both homozygous and heterozygous individuals. No cloning steps are needed and the immobilization and separation of the DNA is accomplished using magnetic beads.
Syvänen
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Orion Corporation, Orion Pharmaceutica Biotechnology, Helsinki, Finland.
Hultman
T
T
Aalto-Setälä
K
K
Söderlund
H
H
Uhlén
M
M
D001057:Apolipoproteins E / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic / Q000235:genetics*; D012371:Robotics; D016415:Sequence Alignment / Q000379:methods*
D001057:Apolipoproteins E; D015345:Oligonucleotide Probes