title: Improvements to bead-based oligonucleotide ligation SNP genotyping assays.
abstract: We describe a bead-based, multiplexed, oligonucleotide ligation assay (OLA) performed on the Luminex flow cytometer. Differences between this method and those previously reported include the use of far fewer beads and the use of a universal oligonucleotide for signal detection. These innovations serve to significantly reduce the cost of the assay, while maintaining robustness and accuracy. Comparisons are made between the Luminex OLA and both pyrosequencing and direct sequencing. Experiments to assess conversion rates, call rates, and concordance across technical replicates are also presented.
authors.affiliation: Department of Genetics, Rutgers University, Piscataway, NJ 08854, USA. shannon.bruse@mirusbio.com
authors.forename: Shannon
authors.initials: S
authors.lastname: Bruse
authors.forename: Michael
authors.initials: M
authors.lastname: Moreau
authors.forename: Marco
authors.initials: M
authors.lastname: Azaro
authors.forename: Ray
authors.initials: R
authors.lastname: Zimmerman
authors.forename: Linda
authors.initials: L
authors.lastname: Brzustowicz
mesh_terms: D000483:Alleles; D001681:Biological Assay / Q000191:economics / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism; D005821:Genetic Techniques; D015894:Genome, Human; D005838:Genotype; D006801:Humans; D008863:Microspheres; D009841:Oligonucleotides / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide
chemical_list: D005456:Fluorescent Dyes; D009841:Oligonucleotides