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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.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1175827
Patent Number(s):
7074565
Application Number:
10/439,507
Assignee:
The Regents of the University of California (Los Alamos, NM)
Patent Classifications (CPCs):
C - CHEMISTRY C07 - ORGANIC CHEMISTRY C07H - SUGARS
Y - NEW / CROSS SECTIONAL TECHNOLOGIES Y10 - TECHNICAL SUBJECTS COVERED BY FORMER USPC Y10S - TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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}
}

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Works referenced in this record:

An agent cleaving glucose-derived protein crosslinks in vitro and in vivo
journal, July 1996


Relief of amplification inhibition in PCR with bovine serum albumin or T4 gene 32 protein.
journal, January 1996


Detection of low numbers of bacterial cells in soils and sediments by polymerase chain reaction.
journal, January 1992


Rapid DNA extraction protocol from soil for polymerase chain reaction-mediated amplification
journal, January 1993


An effective method to extract DNA from environmental samples for polymerase chain reaction amplification and DNA fingerprint analysis
journal, November 1994


Distribution and abundance of Gram-positive bacteria in the environment: development of a group-specific probe
journal, April 2001


DNA recovery from soils of diverse composition.
journal, January 1996


Small-Scale DNA Sample Preparation Method for Field PCR Detection of Microbial Cells and Spores in Soil
journal, July 1998


Rapid method for separation of bacterial DNA from humic substances in sediments for polymerase chain reaction.
journal, January 1992