UNIT 4.15 TaqMan Real Time–Polymerase Chain Reaction Methods for Determination of Nucleotide Polymorphisms in Human N-Acetyltransferase-1 (NAT1) and -2 (NAT2)
Published Online: 1 JAN 2005
Copyright © 2004 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Toxicology
How to Cite
Hein, D. W. and Doll, M. A. 2005. TaqMan Real Time–Polymerase Chain Reaction Methods for Determination of Nucleotide Polymorphisms in Human N-Acetyltransferase-1 (NAT1) and -2 (NAT2). Current Protocols in Toxicology. 22:4.15:4.15.1–4.15.11.
- Published Online: 1 JAN 2005
- Published Print: NOV 2004
N-acetyltransferase 1 (NAT1) and N-acetyltransferase 2 (NAT2) exhibit allelic variation and genetic polymorphism associated with increased susceptibility towards drug toxicity and environmental disease. TaqMan allelic discrimination methods are described to rapidly determine NAT1 and NAT2 genotypes. The SNPs selected for NAT1 genotype determinations are: C97T (R33Stop), C190T (R64W), G445A (V149I), C559T (R187Stop), G560A (R187Q), A752T (D251V), T1088A (3′UTR), and C1095A (3′UTR). The SNPs selected for NAT2 genotyping determinations are: G191A (R64Q), C282T (silent), T341C (I114T), C481T (silent), G590A (R197Q), A803G (K268R), and G857A (G286T). All NAT2 and NAT1 alleles, except very rare ones, are detected with these assays. Major advantages of the methods described in this unit are that they do not require post-PCR processing (such as enzyme digestion) or the use of radioactivity. Since the methods amplify relatively small segments of NAT1 or NAT2, they are effective for human DNA samples derived from buccal cells or paraffin-embedded tissues.
- acetylator genotype;
- allele discrimination