Simple non-radioactive methods of analysis of polymorphic markers flanking human apolipoprotein C-III gene.
We describe two rapid non-radioactive methods for the analysis of polymorphic markers in the flanking region of the human apolipoprotein CIII gene. The polymorphic markers comprise previously described variable sites located upstream from the coding region of the gene (C-641-->A, G-630-->A, T-625-->deletion, C-482-->T, and T-455-->C) and a polymorphic SstI/SacI site in the apoC-III 3' untranslated region. The first method is allele-specific amplification (ASA) with primers complementary to the normal ("wild-type") allele or to the variable ("mutant") allele at their 3' ends. The other is allele-specific oligonucleotide hybridization (ASO hybridization) with pairs of probes labeled by digoxigenin. Comparison with sequencing data showed that both methods are reliable for polymorphism analysis.
Surguchov
A P
AP
Department of Medicine, Baylor College of Medicine, Houston, TX 77030, USA.
Liu
S B
SB
Patsch
W
W
D053305:Apolipoprotein C-III; D001056:Apolipoproteins C / Q000235:genetics*; D001483:Base Sequence; D005796:Genes; D005819:Genetic Markers; D005821:Genetic Techniques; D005838:Genotype; D006801:Humans; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000235:genetics; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic
D053305:Apolipoprotein C-III; D001056:Apolipoproteins C; D005819:Genetic Markers; D015335:Molecular Probes; D015345:Oligonucleotide Probes