GENOTYPING WITH MINOR-GROOVE-BINDER-CONTAINING PROBES.
APPLICANT'S DESCRIPTION: DNA fingerprinting, or profiling, is widely used for
paternity cases and criminal investigations (forensics). Current methodologies
are cumbersome, expensive and often fail to produce interpretable results. A
minor groove binder (MGB) has been developed that greatly enhances the ability
of oligonucleotide probes to discriminate single nucleotide polymorphisms
(SNPs) in the 5' nuclease (Taqman (registered), real time PCR, fluorescence
quench release) assay. The goal is to develop a kit for rapid high throughput
DNA fingerprinting using probes that contain MGB's and other proprietary
modifications. The fingerprint is derived from a panel of 23 diallelic
polymorphisms. The probability that two unrelated individuals would have
matching fingerprints is approximately 1 in 10 (exp) 8. In the Phase I effort,
PCR primers and probes for the 23 loci were optimized and their function was
demonstrated in 5' nuclease assays using a set of known DNA samples. The goal
of phase II is to develop a commercial prototype with increased throughput and
reduced cost. Application of four spectrally separated reporter fluorophores
and a novel quencher will enable multiplexed assays. Standard affordable
instruments will replace expensive instruments specialized for 5' nuclease
assays. The technologies developed herein can be readily applied to assays of
any SNP's. Huge potential markets exist for these technologies in clinical
diagnostics and pharmacogenomics.
PROPOSED COMMERCIAL APPLICATION:
The commercial prototype presented here is a kit for human identification in paternity testing.
The technologies developed herein for single nucleotide polymorphism (SNP) screening could
have wide market potentials in a variety of applications including clinical diagnostics and
Small Business Information at Submission:
Principal Investigator:Michael J. Singer
EPOCH BIOSCIENCES, INC.
21720 23RD DR SE, #150 BOTHELL, WA 98021
Number of Employees: