title: Integration of DNA ligation and rolling circle amplification for the homogeneous, end-point detection of single nucleotide polymorphisms.
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.
authors.affiliation: Amersham Biosciences UK Ltd, The Grove Centre, AL16, White Lion Road, Amersham, Buckinghamshire HP7 9LL, UK. judith.pickering@uk.amershambiosciences.com
authors.forename: Judith
authors.initials: J
authors.lastname: Pickering
authors.forename: Anona
authors.initials: A
authors.lastname: Bamford
authors.forename: Varsha
authors.initials: V
authors.lastname: Godbole
authors.forename: Jackie
authors.initials: J
authors.lastname: Briggs
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Scozzafava
authors.forename: Phyllida
authors.initials: P
authors.lastname: Roe
authors.forename: Claire
authors.initials: C
authors.lastname: Wheeler
authors.forename: Firman
authors.initials: F
authors.lastname: Ghouze
authors.forename: Sarah
authors.initials: S
authors.lastname: Cuss
mesh_terms: D000483:Alleles; D001331:Automation; D004247:DNA / Q000737:chemistry; D017931:DNA Primers; D004270:DNA, Circular / Q000737:chemistry; D023381:Endpoint Determination; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry; D020641:Polymorphism, Single Nucleotide; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D017931:DNA Primers; D004270:DNA, Circular; D015345:Oligonucleotide Probes; D004247:DNA