Single-track sequencing for genotyping of multiple SNPs in the N-acetyltransferase 1 (NAT1) gene

单轨测序用于 N-乙酰转移酶 1 (NAT1) 基因中多个 SNP 的基因分型

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作者:Pavel Soucek, Camilla Furu Skjelbred, Marit Svendsen, Tom Kristensen, Elin H Kure, Vessela N Kristensen

Background

Fast, cheap and reliable

Conclusions

The method could be of great help to scientists in the field of molecular epidemiology of screening of large populations for known informative biomarkers of susceptibility, such as NAT1.

Results

A strategy allowing to address simultaneously 24 various genetic variants in the NAT1 gene using the single sequencing reaction method on the same PCR product is described. A modified automated DNA sequencing using only one of the sequence terminators was used to genotype PCR products in single-track sequencing reactions of NAT1 and was shown to be universal for both DNA sequencing using labeled primers and labeled nucleotides. By this method we detected known SNPs at site T640G, which confers the NAT1*11 allele with frequency of 0.036, further T1088A and C1095A with frequency of 0.172 and 0.188, respectively and a deletion of TAATAATAA in the poly A signal area with a frequency 0.031. All observed frequencies were in Hardy Weinberg equilibrium and comparable to those in Caucasian population. The single-track signatures of the variant genotypes were verified on samples previously genotyped by RLFP. Conclusions: The method could be of great help to scientists in the field of molecular epidemiology of screening of large populations for known informative biomarkers of susceptibility, such as NAT1.

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