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Nucleic Acids Research, 2002, Vol. 30, No. 12 e60
© 2002 Oxford University Press

Integration of DNA ligation and rolling circle amplification for the homogeneous, end-point detection of single nucleotide polymorphisms

Judith Pickering*, Anona Bamford, Varsha Godbole, Jackie Briggs, Giuseppe Scozzafava, Phyllida Roe, Claire Wheeler, Firman Ghouze and Sarah Cuss

Amersham Biosciences UK Ltd, The Grove Centre, AL16, White Lion Road, Amersham, Buckinghamshire HP7 9LL, UK

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 AmplifluorTM 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.

* To whom correspondence should be addressed. Tel: +44 1494 543984; Fax: +44 1494 543627; Email: judith.pickering{at}uk.amershambiosciences.com


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