Detection of single nucleotide polymorphism in human IL‐4 receptor by PCR amplification of specific alleles

A key aspect of genomic research in the “post‐genome era” is to associate sequence variations with heritable phenotypes. The most common variations in the human genome are single nucleotide polymorphisms (SNPs) that occur approximately once in every 500 to 1,000 bases. Although analyzing the phenotypic outcome of these SNPs is crucial to facilitate large‐scale association studies of genetic diseases, detection of SNPs from an extended number of human DNA samples is often difficult, labor‐intensive and time‐consuming. Recent development in SNP detection methods using DNA microarrays and mass spectrophotometry has allowed automated high throughput analyses, but such equipments are not accessible to many scientists. In this study, we demonstrate that a simple PCR‐based method using primers with a mismatched base at the 3'‐end provides a fast and easy tool to identify known SNPs from human genomic DNA in a regular molecular biology laboratory. Results from this PCR amplification of specific alleles (PASA) analysis efficiently and accurately typed the Q576R polymorphism of human IL4 receptor from the genomic DNAs of 29 Koreans, including 9 samples whose genotype could not be discerned by the conventional PCR‐SSCP (single strand conformation polymorphism) method. Given the increasing attention to disease‐associated polymorphisms in genomic research, this alternative technique will be very useful to identify SNPs in large‐scale population studies.

Key words:

• Human IL4 receptor
• Single nucleotide polymorphism
• PASA

Notes

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