NEW APPROACH TO RAPID AND SENSITIVE DNA QUANTITATION

Information

  • Research Project
  • 2536724
  • ApplicationId
    2536724
  • Core Project Number
    R43GM057092
  • Full Project Number
    1R43GM057092-01
  • Serial Number
    57092
  • FOA Number
  • Sub Project Id
  • Project Start Date
    9/30/1997 - 26 years ago
  • Project End Date
    9/30/1998 - 25 years ago
  • Program Officer Name
  • Budget Start Date
    9/30/1997 - 26 years ago
  • Budget End Date
    9/30/1998 - 25 years ago
  • Fiscal Year
    1997
  • Support Year
    1
  • Suffix
  • Award Notice Date
    9/27/1997 - 26 years ago
Organizations

NEW APPROACH TO RAPID AND SENSITIVE DNA QUANTITATION

The project will examine the potential of producing an extremely sensitive DNA measurement system based upon the transformation of the nucleotides in DNA to mononucleotide triphosphates followed by detection of the triphosphates using Luciferase. Two different strategies will be explored for the formation of nucleotide triphosphates from DNA: direct generation of the triphosphates by reversing the polymerization reaction of polymerases, and; digestion of the polymer with DNAses followed by transformation of the nucleotide monophosphates to triphosphates using coupled enzyme systems. Phase I of the work will be used to: a) determine the ability of various enzymes to carry out transformations of nucleotides at very low levels of substrate; b) purify in-house the required enzymes necessary for Phase II and; c) determine the detection limit for the combined system. The best systems will then be optimized during Phase II to achieve a detection limit below 100pgrams of DNA and the effect of many agents used to stabilize and purify protein will be determined. During Phase Iii, an automated system to perform the assay will be developed and software for automated data acquisition will be created. PROPOSED COMMERCIAL APPLICATION: The primary market for a DNA Quantitation Kit is the biological pharmaceutical industry. There is a strong F.D.A. mandated need to control the level of DNA contamination to a level less than 100pg. The success of this proposed research will yield one of the most sensitive methods for detecting trace levels of DNA contamination.

IC Name
NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
  • Activity
    R43
  • Administering IC
    GM
  • Application Type
    1
  • Direct Cost Amount
  • Indirect Cost Amount
  • Total Cost
  • Sub Project Total Cost
  • ARRA Funded
  • CFDA Code
    821
  • Ed Inst. Type
  • Funding ICs
  • Funding Mechanism
  • Study Section
    ZRG2
  • Study Section Name
  • Organization Name
    PROMEGA CORPORATION
  • Organization Department
  • Organization DUNS
  • Organization City
    MADISON
  • Organization State
    WI
  • Organization Country
    UNITED STATES
  • Organization Zip Code
    537115399
  • Organization District
    UNITED STATES