Simultaneous genotyping of single nucleotide polymorphisms in the IL-6, IL-10, TNFα and TNFβ genes
Journal
Tissue Antigens
Journal Volume
59
Journal Issue
4
Pages
280-286
Date Issued
2002
Author(s)
Lin M.-T.
Singleton K.
Martin E.G.
Yen A.-H.
Chuang S.-M.
Martin P.J.
Hansen J.A.
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.
SDGs
Other Subjects
endonuclease; interleukin 10; interleukin 6; lymphotoxin; tumor necrosis factor alpha; cytokine; interleukin 10; interleukin 6; lymphotoxin; primer DNA; tumor necrosis factor alpha; article; base mispairing; gene amplification; gene function; genetic susceptibility; genotype; human; nucleotide sequence; polymerase chain reaction; priority journal; restriction fragment length polymorphism; restriction site; single nucleotide polymorphism; genetics; genotype; methodology; polymerase chain reaction; restriction fragment length polymorphism; Cytokines; DNA Primers; Genotype; Human; Interleukin-10; Interleukin-6; Lymphotoxin; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Polymorphism, Single Nucleotide; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.; Tumor Necrosis Factor; DNA Primers; Genotype; Humans; Interleukin-10; Interleukin-6; Lymphotoxin-alpha; Tumor Necrosis Factor-alpha
Type
journal article