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DNA Repair Proficient PCR

Award Information

Agency:
Department of Health and Human Services
Branch:
N/A
Award ID:
93865
Program Year/Program:
2009 / SBIR
Agency Tracking Number:
GM088954
Solicitation Year:
N/A
Solicitation Topic Code:
N/A
Solicitation Number:
N/A
Small Business Information
LUCIGEN CORPORATION
2905 Parmenter St. MIDDLETON, WI -
View profile »
Woman-Owned: No
Minority-Owned: No
HUBZone-Owned: No
 
Phase 1
Fiscal Year: 2009
Title: DNA Repair Proficient PCR
Agency: HHS
Contract: 1R43GM088954-01
Award Amount: $99,050.00
 

Abstract:

DESCRIPTION (provided by applicant): The Polymerase Chain Reaction (PCR) is one of the most widely used techniques in molecular biology. Unfortunately, thermocycling at elevated temperatures (60o to 95oC) causes extensive thermal damage to DNA; in particu lar, A+G depurination. In vivo, apurinic sites are repaired by AP lyase enzymes, which cleave DNA to form elongatable 3'OH ends. However, in vitro PCR reactions generally lack DNA repair enzymes. Since apurinic sites are not removed, DNA synthesis terminat es. In this Phase I Research grant application, enzymology is described for DNA repair- proficient PCR. A novel thermostable enzyme mixture will be developed, using recombinant Thermotoga maritima AP lyase, PyroPhage 3173 polymerase, and an engineered pyro phosphatase. In DNA repair-proficient PCR, sites of A+G depurination are enzymatically hydrolysed to 3'OH ends during thermocycling, so that thermally damaged DNA strands are continually converted into elongatable molecules. As a result, it is hypothesized that the yield, sensitivity, and amplicon length of PCR can be increased. PDF Creator - PDF4Free v2.0 http://www.pdf4free.com PUBLIC HEALTH RELEVANCE: The Polymerase Chain Reaction (PCR) is one of the most widely used techniques in molecular medici ne for the diagnosis of heritable and infectious diseases. However, the high-temperature reaction conditions employed during PCR result in thermal damage to DNA. The proposed research describes a repair-proficient enzyme mixture to improve the reaction yie ld, sensitivity, and throughput of DNA-based diagnostics. PDF Creator - PDF4Free v2.0 http://www.pdf4free.com

Principal Investigator:

Jeffrey Williams
JWILLIAMS@LUCIGEN.COM

Business Contact:

David Mead
dmead@lucigen.com
Small Business Information at Submission:

LUCIGEN CORPORATION
2120 W. GREENVIEW DR. SUITE 9 MIDDLETON, WI 53562

EIN/Tax ID: 391923306
DUNS: N/A
Number of Employees: N/A
Woman-Owned: No
Minority-Owned: No
HUBZone-Owned: No