Premium
Multiple sample amplification and genotyping integrated on a single electronic microarray
Author(s) -
Huang Ying,
Shirajian Jack,
Schroder Astrid,
Yao Zuxu,
Summers Tamara,
Hodko Dalibor,
Sosnowski Ron
Publication year - 2004
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200406075
Subject(s) - genotyping , multiple displacement amplification , microbiology and biotechnology , dna microarray , biology , snp genotyping , genomic dna , molecular inversion probe , locus (genetics) , genotype , primer (cosmetics) , single nucleotide polymorphism , locked nucleic acid , dna , genetics , oligonucleotide , polymerase chain reaction , gene , dna extraction , chemistry , gene expression , organic chemistry
We report a novel method that allows simultaneous in situ amplification and then genotyping of single nucleotide polymorphism (SNP) for multiple samples on a single electronic microarray. The locus coding for one of the common inherited thrombosis risk factors, Factor V Leiden (FVL), was chosen as a model system for SNP analysis. This method combines strand displacement amplification (SDA) with electrophoretic movement and concentration of DNA on electronic microarrays to provide a single platform for DNA amplification and analysis. The method includes: electronic anchoring of allele‐specific SDA amplifiable primers (APs) and a nonamplifiable primer (NAP) to different electrodes, electronic hybridization of genomic DNA from different samples to those primers, in situ amplification of target DNA, and genotyping of FVL. Compared to previous anchored SDA methods, the addition of a NAP improves detection signals by at least 20‐fold. The sensitivity of this method is dependent on the amplification time. Using this method, nine different genomic DNA samples with known FVL genotypes were amplified and correctly genotyped on a single electronic microarray without any contamination between samples. The present method could streamline development of nucleic acid‐based assays in applications of molecular diagnostic, point‐of‐care testing, and forensic detection, which often require the capability to analyze multiple samples efficiently.