TY  - JOUR
A1  - Mein, Charles A.
A1  - Barratt, Bryan J.
A1  - Dunn, Michael G.
A1  - Siegmund, Thorsten
A1  - Smith, Annabel N.
A1  - Esposito, Laura
A1  - Nutland, Sarah
A1  - Stevens, Helen E.
A1  - Wilson, Amanda J.
A1  - Phillips, Michael S.
A1  - Jarvis, Nancy
A1  - Law, Scott
A1  - de Arruda, Monika
A1  - Todd, John A.
T1  - Evaluation of Single Nucleotide Polymorphism Typing with Invader on PCR Amplicons and Its Automation
Y1  - 2000/03/01 
JF  - Genome Research 
JO  - Genome Research 
SP  - 330 
EP  - 343 
DO  - 10.1101/gr.10.3.330 
VL  - 10 
IS  - 3 
UR  - http://genome.cshlp.org/content/10/3/330.abstract 
N2  - Large-scale pharmacogenetics and complex disease association studies will require typing of thousands of single-nucleotide polymorphisms (SNPs) in thousands of individuals. Such projects would benefit from a genotyping system with accuracy >99% and a failure rate <5% on a simple, reliable, and flexible platform. However, such a system is not yet available for routine laboratory use. We have evaluated a modification of the previously reported Invader SNP-typing chemistry for use in a genotyping laboratory and tested its automation. The Invader technology uses a Flap Endonuclease for allele discrimination and a universal fluorescence resonance energy transfer (FRET) reporter system. Three hundred and eighty-four individuals were genotyped across a panel of 36 SNPs and one insertion/deletion polymorphism with Invader assays using PCR product as template, a total of 14,208 genotypes. An average failure rate of 2.3% was recorded, mostly associated with PCR failure, and the typing was 99.2% accurate when compared with genotypes generated with established techniques. An average signal-to-noise ratio (9:1) was obtained. The high degree of discrimination for single base changes, coupled with homogeneous format, has allowed us to deploy liquid handling robots in a 384-well microtitre plate format and an automated end-point capture of fluorescent signal. Simple semiautomated data interpretation allows the generation of ∼25,000 genotypes per person per week, which is 10-fold greater than gel-based SNP typing and microsatellite typing in our laboratory. Savings on labor costs are considerable. We conclude that Invader chemistry using PCR products as template represents a useful technology for typing large numbers of SNPs rapidly and efficiently. 
ER  -