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High Resolution Fluorescent Melting Curve Analysis

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: PHS2001-2
Agency Tracking Number: 2R42GM060063-02
Amount: $0.00
Phase: Phase I
Program: STTR
Solicitation Topic Code: N/A
Solicitation Number: N/A
Timeline
Solicitation Year: N/A
Award Year: 2001
Award Start Date (Proposal Award Date): N/A
Award End Date (Contract End Date): N/A
Small Business Information
390 WAKARA WAY SALT LAKE CITY, UT 84108-1214
SALT LAKE CITY, UT 84108
United States
DUNS: N/A
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 PHILIP BERNARD
 (801) 581-4213
 PHIL.BERNARD@PATH.UTAH.EDU
Business Contact
 BOTOSAN, MONTY
Phone: (801) 582-5138
Email: MONTY@IDAHOTEC.COM
Research Institution
N/A
Abstract

DESCRIPTION (Applicant's abstract): Our long-term goal is to advance
instrumentation and techniques for rapid, automated and sensitive variant
analysis of nucleic acids. The LightCyclerTM has been our homogenous "closed
system" platform for genotyping. Fluorescent hybridization probes are included
at the beginning of the reaction and are used for realtime PCR monitoring and
genotyping. After rapid cycle PCR, the reaction is slowly heated (0.1 degree
C/s) and products are identified by unique probe/target melting temperatures.
The entire assay is performed from genomic DNA within 20 min and without any
sample manipulation. Fluorescent melting curve analysis is a useful and popular
method for repetitive genotyping of established mutations/polymorphisms. In
phase I, we built and tested a new "high-resolution" instrument for melting
curve analysis. The high-resolution melting instrument has 10-fold less
measurement error than the LightCyclerTM providing increased sensitivity for
mutation detection and utility in mutation scanning. In phase II, we plan to
integrate the "low noise" components of the high-resolution instrument into a
modified LightCyclerTM to allow both rapid amplification and smooth melting
curve genotyping. Mathematical software algorithms will be written for
sensitive variant detection and flagging unknown mutant sequences. The utility
of the new instrument and algorithms will be demonstrated by homogenously
scanning for mutations within critical exons of the p53 gene.
PROPOSED COMMERCIAL APPLICATION:
We propose developing instrumentation and analysis software that will allow homogenous
mutation scanning for the first time. The system will be used for scanning the p53 tumor
suppressor gene, which is mutated in almost all cancers at the time of invasive growth.
The instrumentation and the assay will be commercially available for continued research
on p53 and to provide a clinical test for the prognosis and treatment of cancer. The
general method can be applied to other genes, as well.

* Information listed above is at the time of submission. *

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