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Preparation of DNA-containing extract for PCR amplification

United States Patent

July 11, 2006
View the Complete Patent at the US Patent & Trademark Office
Los Alamos National Laboratory - Visit the Technology Transfer Division Website
Environmental samples typically include impurities that interfere with PCR amplification and DNA quantitation. Samples of soil, river water, and aerosol were taken from the environment and added to an aqueous buffer (with or without detergent). Cells from the sample are lysed, releasing their DNA into the buffer. After removing insoluble cell components, the remaining soluble DNA-containing extract is treated with N-phenacylthiazolium bromide, which causes rapid precipitation of impurities. Centrifugation provides a supernatant that can be used or diluted for PCR amplification of DNA, or further purified. The method may provide a DNA-containing extract sufficiently pure for PCR amplification within 5 10 minutes.
Dunbar; John M. (Sante Fe, NM), Kuske; Cheryl R. (Los Alamos, NM)
The Regents of the University of California (Los Alamos, NM)
10/ 439,507
May 15, 2003
STATEMENT REGARDING FEDERAL RIGHTS This invention was made with government support under Contract No. W-7405-ENG-36 awarded by the U.S. Department of Energy. The government has certain rights in the invention.