Journal
JOURNAL OF BIOTECHNOLOGY
Volume 135, Issue 2, Pages 157-160Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2008.03.011
Keywords
single nucleotide polymorphism (SNP) typing; allele-specific PCR; L-DNA; surface plasmon resonance (SPR) imaging
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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. (c) 2008 Elsevier B.V. All rights reserved.
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