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  • Title: Genotyping of multiple single nucleotide polymorphisms with hyperbranched rolling circle amplification and microarray.
    Author: Li X, Luo J, Xiao P, Shi X, Tang C, Lu Z.
    Journal: Clin Chim Acta; 2009 Jan; 399(1-2):40-4. PubMed ID: 18775688.
    Abstract:
    BACKGROUND: The emerging role of single nucleotide polymorphisms (SNPs) in clinical diagnostics and studies has created a need for simple and high-throughput genotyping methods. Previously, we developed a 3-dimensional polyacrylamide gel-based microarray (3-D microarray) of PCR-product. This method can detect single SNP locus from multiple DNA samples on one chip. METHODS: Hyperbranched rolling circle amplification (HRCA) was used to recognize different SNP loci and amplify the fragments from genomic samples. Different HRCA products were used to fabricate the 3-D microarray, and dual-color fluorescent probes were used to detect signals. RESULTS: This assay was applied to genotype 2 SNP loci from a set of 6 genomic DNA samples on one chip. Universal acryl-modified primer and one pair of dual-color fluorescent probes were used for all sample detection to reduce the cost. We demonstrate that this assay can detect 10 ng genomic DNA. CONCLUSIONS: Combination of HRCA and 3-D microarray allows parallel discrimination of different alleles from different samples on a single chip. It is a feasible method for high-throughput mutation analysis and disease diagnosis.
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