Simultaneous genotyping of single nucleotide polymorphisms in the IL-6, IL-10, TNFα and TNFβ genes

L. H. Tseng, P. J. Chen, M. T. Lin, K. Singleton, E. G. Martin, A. H. Yen, S. M. Chuang, P. J. Martin, J. A. Hansen

Research output: Contribution to journalArticlepeer-review

Abstract

Single nucleotide polymorphisms (SNP) in the human IL-6, IL-10, TNFα and TNFβ genes have been associated with gene function and susceptibility to disease. In this study, primers containing mismatches at 1-3 nucleotide positions were designed to incorporate a new restriction site recognized by endonucleases AlwNI, BcgI, BglI, BsaBI, BslI, BstXI, EcoNI or XcmI for genotyping SNPs in the IL-6 gene (position - 174), IL-10 gene (positions -592 and -1082), TNFα gene (positions -238, -308 and -863) and TNFβ gene (position + 249) by mismatched polymerase chain reaction and restriction fragment length polymorphism (PCR/RFLP). Our results show that appropriately designed BslI-based mismatched PCRfRFLP assays can be successfully used to determine the genotypes for approximately 40% of SNPs. The mismatched PCR strategy can be coupled with multiplex-amplification to enable simple and rapid determination of several SNP genotypes in a single reaction.

Original languageEnglish (US)
Pages (from-to)280-286
Number of pages7
JournalTissue Antigens
Volume59
Issue number4
DOIs
StatePublished - 2002
Externally publishedYes

Keywords

  • Bsll
  • Multiplex
  • PCR
  • RFLP

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Biochemistry
  • Genetics

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