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HOMOGENEOUS MUTATION SCANNING

Award Number: R42GM072419
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: HEALTH SERVICES
PERIOD OF PERFORMANCE START DATE: 08/01/2004
PERIOD OF PERFORMANCE END DATE: 01/31/2007

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HOMOGENEOUS MUTATION SCANNING - DESCRIPTION (provided by applicant): Our objective is to provide a homogeneous, closed-tube system for DNA mutation scanning on a high-throughput platform. For many complex genetic diseases, it is very difficult and expensive to screen for all possible mutations that may cause the disease. We propose a relatively simple solution. Certain dyes are compatible with PCR and can detect sequence alterations present in one copy of DNA by simple melting analysis after amplification. The dye is added before PCR, and optionally followed in real-time during amplification. Heterozygotes are easily identified after a 5 min. high-resolution melting curve after PCR. Specific aims for Phase I of this Fast Track proposal include: 1. Identify and/or synthesize scanning dyes that can be used with standard fluorescein optics. 2. Demonstrate that heterozyotes can be identified by high-resolution melting on a 384-well platform. 3. Achieve 90% sensitivity and specificity for single base change heterozygote detection in PCR products up to 300 bps in length. Our currently manufactured 384-well, low-resolution genotyping instrument will be modified for high-resolution scanning. A robust DNA scanning dye for use with standard fluorescein optics will be identified and/or synthesized for this instrument and potentially others. A comprehensive screen of single base mismatches in engineered plasmids will be used to establish the sensitivity and specificity of the system, and provide the criteria for Phase II funding. Advantages of high-resolution melting analysis for mutation scanning include speed (< 5 min), homogeneous design (no need for automation), closed-tube analysis (no amplicon contamination risk), and sample disposition (immediately available for sequencing if 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	1/19/2006	NON-COMPETING CONTINUATION	$375,000
														Subtotal = $375,000

Issue Date FY: 2005 ( Subtotal = $375,000 )
2005	2005	IDAHO TECHNOLOGY	149 CHESTNUT STREET	IDAHO FALLS	ID	83402	BONNEVILLE	USA	Biomedical Research and Research Training	000	2	1/31/2005	EXTENSION WITH OR WITHOUT FUNDS	$375,000
														Subtotal = $375,000

Issue Date FY: 2004 ( Subtotal = $100,000 )
2004	2004	IDAHO TECHNOLOGY	149 CHESTNUT STREET	IDAHO FALLS	ID	83402	BONNEVILLE	USA	Biomedical Research and Research Training	000	1	7/23/2004	NEW	$100,000
														Subtotal = $100,000

Grand Total All Awards = $850,000

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HOMOGENEOUS MUTATION SCANNING

Award Number: R42GM072419

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: HEALTH SERVICES

PERIOD OF PERFORMANCE START DATE: 08/01/2004

PERIOD OF PERFORMANCE END DATE: 01/31/2007

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