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Allele-Specific Real-Time Polymerase Chain Reaction as a Tool for Urate Transporter 1 Mutation Detection

  • Juliet O. Makanga
  • Antonius Christianto
  • Tetsuya Inazu
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1275)

Abstract

Allele-specific polymerase chain reaction (ASPCR) method has long been applied for the detection of nucleotide variations and genotyping, which are detected by the presence or absence of DNA amplification PCR products. Recently, Real-Time PCR genotyping has fast developed and offered a rapid method of detecting mutations without the need of gel electrophoresis as with ASPCR. Here, we describe an easy and rapid touchdown real-time PCR method for the detection of nucleotide variations. Using our method we successfully detect two main mutations in human urate transporter 1 (SLC22A12), W258X and R90H, and validate the results. The method can potentially be applied to genotype of various other nucleotide variations.

Key words

Allele-specific real-time polymerase chain reaction Urate transporter Genotyping 

References

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Juliet O. Makanga
    • 1
  • Antonius Christianto
    • 1
  • Tetsuya Inazu
    • 1
  1. 1.Laboratory of Functional Genomics, Department of Pharmacy, College of Pharmaceutical SciencesRitsumeikan UniversityKusatsu CityJapan

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