SIMPLE NONRADIOACTIVE METHODS OF ANALYSIS OF POLYMORPHIC MARKERS FLANKING HUMAN APOLIPOPROTEIN C-III GENE

We describe two rapid non-radioactive methods for the analysis of poly morphic markers in the flanking region of the human apolipoprotein CII I 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. T he first method is allele-specific amplification (ASA) with primers co mplementary to the normal (''wild-type'') allele or to the variable (' 'mutant'') allele at their 3' ends. The other is allele-specific oligo nucleotide hybridization (ASO hybridization) with pairs of probes labe led by digoxigenin. Comparison with sequencing data showed that both m ethods are reliable for polymorphism analysis.