Genotyping by allele-specific L-DNA-tagged PCR.
We have developed a novel method for typing single-nucleotide polymorphisms (SNPs) that can be applicable to rapid screening. The method involves the fusion of two PCR techniques, allele-specific PCR (AS-PCR) and L-DNA-tagged PCR (LT-PCR), which enables us to label PCR products with sequence-defined tags of mirror-image DNA (L-DNA). PCR products were applied without any purification or denaturation steps to gold surfaces where complementary single-stranded L-DNA was immobilized, and the products were detected with surface plasmon resonance (SPR) imaging. We were able to clearly discriminate 3 genotypes at position 2677 of the MDR1 gene (G/G-homozygote, G/T-heterozygote, and T/T-homozygote) by comparing SPR difference images.
Hayashi
Gosuke
G
Department of Regulatory Bioorganic Chemistry, The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki 567-0047, Japan.
Hagihara
Masaki
M
Nakatani
Kazuhiko
K
D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics; D000483:Alleles; D004247:DNA / Q000235:genetics*; D005838:Genotype; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D008957:Models, Genetic; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D020349:Surface Plasmon Resonance
C513055:ABCB1 protein, human; D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D004247:DNA