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TaqMan real time-polymerase chain reaction methods for determination of nucleotide polymorphisms in human N-acetyltransferase-1 (NAT1) and -2 (NAT2).

Authors
  • 1
  • 1 University of Louisville School of Medicine, Louisville, KY, USA.
Type
Published Article
Journal
Current protocols in toxicology
1934-9262
Publication Date
Volume
Chapter 4
Identifiers
DOI: 10.1002/0471140856.tx0415s22
PMID: 23045122
Source
Medline
License
Unknown

Abstract

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: C(97)T (R(33)Stop), C(190)T (R(64)W), G(445)A (V(149)I), C(559)T (R(187)Stop), G(560)A (R(187)Q), A(752)T (D(251)V), T(1088)A (3'UTR), and C(1095)A (3'UTR). The SNPs selected for NAT2 genotyping determinations are: G(191)A (R(64)Q), C(282)T (silent), T(341)C (I(114)T), C(481)T (silent), G(590)A (R(197)Q), A(803)G (K(268)R), and G(857)A (G(286)T). 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.

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