Improvements to bead-based oligonucleotide ligation SNP genotyping assays.
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.
Bruse
Shannon
S
Department of Genetics, Rutgers University, Piscataway, NJ 08854, USA. shannon.bruse@mirusbio.com
Moreau
Michael
M
Azaro
Marco
M
Zimmerman
Ray
R
Brzustowicz
Linda
L
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
D005456:Fluorescent Dyes; D009841:Oligonucleotides