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A SYSTEM FOR RAPID PCR, MUTATION SCANNING AND GENOTYPING

Award Number: R42GM073396
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: HEALTH SERVICES
PERIOD OF PERFORMANCE START DATE: 03/01/2005
PERIOD OF PERFORMANCE END DATE: 08/31/2007

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A SYSTEM FOR RAPID PCR, MUTATION SCANNING AND GENOTYPING - DESCRIPTION (provided by applicant): Our objective is to provide a closed-tube system for rapid PCR, mutation scanning and genotyping that does not require fluorescently-labeled probes. For many genetic diseases, it is difficult and expensive to screen for all possible mutations that may cause the disease. We propose a relatively simple solution for rapid amplification, scanning and genotyping in a single homogeneous system. Certain DNA dyes are compatible with PCR and can detect mismatches between two copies of DNA by simple melting analysis after amplification. The dye is added before PCR, and heterozygotes are easily identified after a 1-2 min high-resolution melting curve after PCR. Specific genotype confirmation is provided by an unlabeled oligonucleotide probe. The specific aims for Phase I of this Fast Track proposal are: 1. Develop a homogeneous, closed-tube gentotyping method that uses unlabeled oligonucleotides. 2. Prototype a rapid PCR system that integrates high-resolution melting analysis for scanning and genotyping. We will integrate the high-resolution melting capability of our HR-1 instrument into our rapid PCR machine (R.A.P.I.D./LightCycler). After PCR, each tube will be automatically analyzed by high-resolution melting. Simultaneous scanning and genotyping will be demonstrated using special DNA dyes. Advantages of our system includes speed (PCR in 15 min, analysis in 1-2 min/sample), homogeneous design (no need for sample transfer or reagent additions), closed-tube analysis (no amplicon contamination risk), both scanning and genotyping capability, and nondestructive analysis (immediately available for sequencing in the rare case when it is necessary).


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2006 ( Subtotal = $375,000 )
2006	2006	IDAHO TECHNOLOGY	149 CHESTNUT STREET	IDAHO FALLS	ID	83402	BONNEVILLE	USA	Biomedical Research and Research Training	000	3	7/28/2006	NON-COMPETING CONTINUATION	$375,000
														Subtotal = $375,000

Issue Date FY: 2005 ( Subtotal = $475,000 )
2005	2005	IDAHO TECHNOLOGY	149 CHESTNUT STREET	IDAHO FALLS	ID	83402	BONNEVILLE	USA	Biomedical Research and Research Training	000	1	2/28/2005	NEW	$100,000
2005	2005	IDAHO TECHNOLOGY	149 CHESTNUT STREET	IDAHO FALLS	ID	83402	BONNEVILLE	USA	Biomedical Research and Research Training	000	2	8/22/2005	EXTENSION WITH OR WITHOUT FUNDS	$375,000
														Subtotal = $475,000

Grand Total All Awards = $850,000

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A SYSTEM FOR RAPID PCR, MUTATION SCANNING AND GENOTYPING

Award Number: R42GM073396

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: HEALTH SERVICES

PERIOD OF PERFORMANCE START DATE: 03/01/2005

PERIOD OF PERFORMANCE END DATE: 08/31/2007

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