Integration of DNA ligation and rolling circle amplification for the homogeneous,end-point detection of single nucleotide polymorphisms |
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Authors: | Pickering Judith Bamford Anona Godbole Varsha Briggs Jackie Scozzafava Giuseppe Roe Phyllida Wheeler Claire Ghouze Firman Cuss Sarah |
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Institution: | Amersham Biosciences UK Ltd, The Grove Centre, AL16, White Lion Road, Amersham, Buckinghamshire HP7 9LL, UK. judith.pickering@uk.amershambiosciences.com |
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Abstract: | Association studies using common sequence variants or single nucleotide polymorphisms (SNPs) may provide a powerful approach to dissect the genetic inheritance of common complex traits. Such studies necessitate the development of cost-effective, high throughput technologies for scoring SNPs. The method described in this paper for the co-detection of both alleles of a SNP in a single homogeneous reaction combines the specificity of a high fidelity DNA ligation step with the power of rolling circle amplification. The incorporation of Amplifluor™ energy transfer primers enables signal detection in a homogeneous format, making this approach highly amenable to automation. The adaptation of the genotyping method for high throughput screening using conventional liquid handling systems is described. |
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