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REVERSIBLE MAGNESIUM PRECIPITATION FOR HOT-START PCR

Award Information

Agency:
Department of Health and Human Services
Branch:
N/A
Award ID:
61001
Program Year/Program:
2002 / SBIR
Agency Tracking Number:
GM066632
Solicitation Year:
N/A
Solicitation Topic Code:
N/A
Solicitation Number:
N/A
Small Business Information
DNA Polymerase Technology, Inc.
1508 S. Grand Blvd. Saint Louis, MO 63104-1364
View profile »
Woman-Owned: No
Minority-Owned: No
HUBZone-Owned: Yes
 
Phase 1
Fiscal Year: 2002
Title: REVERSIBLE MAGNESIUM PRECIPITATION FOR HOT-START PCR
Agency: HHS
Contract: 1R43GM066632-01
Award Amount: $100,000.00
 

Abstract:

DESCRIPTION (provided by applicant): We have been investigating strategies for novel methods of hot start for PCR. One such strategy involves the sequestering of a critical component of the reaction until the reaction temperature is sufficient to ensure selective annealing of PCR primers. We have investigated reversible precipitates of Mg as a method of sequestering Mg2+ ions, which are a critical component of PCR. Many PCR analyses benefit from or require a hot start, particularly those with rare targets or those with compromised DNA templates, such as diagnostic or forensic analyses. In a normal (room temperature) start, the primers can bind to non-specific sequences, or each other, during reaction setup, which can result in the amplification of competitive or inhibitory products. Current methods of achieving a hot start are tedious, expensive, and/or unreliable. We have identified a precipitated form of Mg that is solublized by thermocycling, but is stable enough at room temperature to inhibit polymerase activity. We have also determined that PCR reactions performed with this reversibly precipitated Mg have the benefit of a hot start. We are proposing to simplify and optimize this reversible precipitate technology for standard PCR and extend it to specialized PCR applications, multiplex PCR, Long and Accurate PCR, and even those applications that use other Mg2+ ion dependent enzymes, such as cDNA synthesis by reverse transcriptases.

Principal Investigator:

Katherine R. Rowlyk
4154289092
KR@KLENTAQ.COM

Business Contact:

Wayne Barnes
3144225201
BARNES@KLENTAQ.COM
Small Business Information at Submission:

DNA POLYMERASE TECHNOLOGY, INC.
DNA POLYMERASE TECHNOLOGY, INC. 1508 S GRAND BLVD ST. LOUIS, MO 63104

EIN/Tax ID: 431824012
DUNS: N/A
Number of Employees: N/A
Woman-Owned: No
Minority-Owned: No
HUBZone-Owned: No