ANYL 146 |
| Combing the easiness for separation of magnetic particles and the high-throughput advantage of microarray, a high-throughput automated approach for single nucleotide polymorphisms (SNPs) genotyping using solid-phase PCR on magnetic composite particles (MCPs) was developed. Biotin-labeled reverse primer was captured by streptavidin magnetic composite particles (SA-MCPs), while PCR products were directly amplified on the surface of SA-MCPs in a 96-well plate. The ssDNA amplified on MCPs was hybridized with a pair of dual-color probes, and then genotype of each sample was simultaneously identified by scanning the microarray printed with the denatured fluorescent probes. All the procedures were performed on an automated workstation equipped with a magnetic separation device. To further make the approach more cost-effective, a pair of universal detecting tags were designed and applied. For the determination of each SNP locus, only 6 cents was needed. The feasibility of the approach was confirmed by the successful interrogation of the M235T polymorphism in angiotensinogen (AGT) gene from 160 samples. With automated operation and microarray based “read-out” technology, this approach promises a novel, simple and labor-saving method for large-scale SNP detection. |
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Nanomaterials in Analytical Chemistry
1:25 PM-5:05 PM, Monday, April 7, 2008 Morial Convention Center -- Rm. 335, Oral
Division of Analytical Chemistry |