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Title: Preparation of DNA-containing extract for PCR amplification

Abstract

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.

Inventors:
;
Issue Date:
Research Org.:
The Regents of the University of California, Los Alamos, NM (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1175827
Patent Number(s):
7,074,565
Application Number:
10/439,507
Assignee:
The Regents of the University of California (Los Alamos, NM)
DOE Contract Number:  
W-7405-ENG-36
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Dunbar, John M., and Kuske, Cheryl R. Preparation of DNA-containing extract for PCR amplification. United States: N. p., 2006. Web.
Dunbar, John M., & Kuske, Cheryl R. Preparation of DNA-containing extract for PCR amplification. United States.
Dunbar, John M., and Kuske, Cheryl R. Tue . "Preparation of DNA-containing extract for PCR amplification". United States. https://www.osti.gov/servlets/purl/1175827.
@article{osti_1175827,
title = {Preparation of DNA-containing extract for PCR amplification},
author = {Dunbar, John M. and Kuske, Cheryl R.},
abstractNote = {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.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2006},
month = {7}
}

Patent:

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