Comparative Analysis of Human DNA Variations by Fluorescence-Based Sequencing of PCR Products

Novel SNPs in a candidate gene, CRTH2, for allergic diseases

“…4 The gene for CRTH2 is located in chromosome 11q12, a region showing linkage to asthma and asthma-associated phenotypes (for review see Hakonarson and Wjst 5 ). Using a sequencing strategy, 6 we have identified three novel SNPs in the CRTH2 gene and confirmed an in silico SNP dbSNP ID:rs545659), following a systematic screen of the exon, exon-intron boundary, 5Ј-and 3Ј-UTR sequences in a Chinese population.…”

“…Genomic DNAs were purified from peripheral blood mononuclear cells of each individual. For discovery of SNP markers, sequencing analyses of the human CRTH2 gene were performed initially using pooled DNA samples from 10 randomly chosen subjects (see also Kwok et al 6 ). Any sequence variants identified in this pool were then genotyped in the entire study population.…”

The molecular basis for genetic polymorphism of human deoxyribonuclease I: identification of the nucleotide substitution that generates the fourth allele

“…The nucleotide/amino acid substitutions underlying the protein polymorphism of DNase I are summarized in Table 1. The results of this study demonstrated that the automated, direct cycle sequencing of double-stranded PCR products provides a highly reproducible method for identification of DNA sequence variations [14]. The isoenzyme ofphenotype 4 is focused to the IEF-PAGE gel region between those of phenotypes 1 and 2.…”

Characterization of publicly available SNPs in the Korean population

“…In the present study, 300 publicly available SNPs were characterized in the Korean population by comparative sequencing approach [Kwok et al, 1994]. Every SNP was tested in four samples: three individual samples and a pool of 21 individuals.…”

“…Allele frequencies of a pool were estimated by comparing the peak heights of the corresponding bases between heterozygous individual and a pooled DNA sample [Kwok et al, 1994]. The normalized peak height from heterozygous individual represents the signal contributed by 50% of the DNA template present that bears that of particular allele.…”