Elsevier

Analytical Biochemistry

Volume 288, Issue 1, 1 January 2001, Pages 106-108
Analytical Biochemistry

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Comprehensive Human NAT2 Genotype Method Using Single Nucleotide Polymorphism-Specific Polymerase Chain Reaction Primers and Fluorogenic Probes

https://doi.org/10.1006/abio.2000.4892Get rights and content

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    Protein concentrations were quantified using Bradford reagent assay, following manufacturer's protocol (Bio-Rad, Hercules, CA). Genotypes for NAT1 and NAT2 were determined as previously described (Doll et al., 2010; Doll and Hein, 2001; Doll and Hein, 2002). Analysis was carried out on the Applied Biosystems StepOne™ Plus (Thermofisher, Carlsbad, CA).

  • Arylamine N-acetyltransferase 2 genotype-dependent N-acetylation of isoniazid in cryopreserved human hepatocytes

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    The nomenclature and functional effects of single nucleotide polymorphisms present in various NAT2 haplotypes and genotypes has been reviewed2,21. NAT2 genotypes and deduced phenotypes were determined as described previously21,22. Controls (no DNA template) were run to ensure that there was no amplification of contaminating DNA.

  • N-Acetyltransferase 2 (NAT2) genetic variation and the susceptibility to noncardiac gastric adenocarcinoma in Taiwan

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    DNA was isolated from frozen white blood cells of the participants. The NAT2 genotype was determined by the SNP-specific polymerase chain reaction (PCR), adopted from the studies by Hein and Doll.31,32 This method can identify the seven most frequent SNPs: 191G > A (rs1801279), 282C > T (rs1041983), 341T > C (rs1801280), 481C > T (rs1799929), 590G > A (rs1799930), 803A > G (rs1208), and 857G > A (rs1799931).

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    Comparison of genotyping between mothers and babies gave an indication that there was linkage disequilibrium across the region in Caucasian populations (Smelt et al., 1998). Although multiple genotyping methods have been described (Doll & Hein, 2001, 2002; Brans et al., 2004), with the economy of DNA sequencing, the use of sequencing across the region has been described in population genetic studies, and the wealth of SNPs at the NAT loci has been useful in anthropological studies. Combining resequencing with the use of advanced bioinformatics tools to analyze data derived from international genome consortia (e.g. HapMap), these studies have been useful in comparing populations of diverse geographical distribution.

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