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Title: Universal nucleic acids sample preparation method for cells, spores and their mixture

Patent ·
OSTI ID:1013054

The present invention relates to a method for extracting nucleic acids from biological samples. More specifically the invention relates to a universal method for extracting nucleic acids from unidentified biological samples. An advantage of the presently invented method is its ability to effectively and efficiently extract nucleic acids from a variety of different cell types including but not limited to prokaryotic or eukaryotic cells and/or recalcitrant organisms (i.e. spores). Unlike prior art methods which are focused on extracting nucleic acids from vegetative cell or spores, the present invention effectively extracts nucleic acids from spores, multiple cell types or mixtures thereof using a single method. Important that the invented method has demonstrated an ability to extract nucleic acids from spores and vegetative bacterial cells with similar levels effectiveness. The invented method employs a multi-step protocol which erodes the cell structure of the biological sample, isolates, labels, fragments nucleic acids and purifies labeled samples from the excess of dye.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE
DOE Contract Number:
W-31-109-ENG-38
Assignee:
The United States of America as represented by the United States Department of Energy (Washington, DC)
Patent Number(s):
7,871,764
Application Number:
12/050,508
OSTI ID:
1013054
Country of Publication:
United States
Language:
English

References (12)

Use of 16S rRNA, 23S rRNA, and gyrB Gene Sequence Analysis To Determine Phylogenetic Relationships of Bacillus cereus Group Microorganisms journal July 2006
Use of 16S rRNA, 23S rRNA, and gyrB Gene Sequence Analysis To Determine Phylogenetic Relationships of Bacillus cereus Group Microorganisms journal August 2004
Dna stability and survival of bacillus subtilis spores in extreme dryness journal June 1995
A solid-phase extraction procedure for DNA purification journal August 1989
Novel Sample Preparation Method for Safe and Rapid Detection of Bacillus anthracis Spores in Environmental Powders and Nasal Swabs journal March 2003
A Method for Extracting RNA from Dormant and Germinating Bacillus subtilis Strain 168 Endospores journal July 2006
A Procedure for Isolating High Quality DNA from Spores of Bacillus subtilis 168 journal February 1980
Continuous Spore Disruption Using Radially Focused, High-Frequency Ultrasound journal August 2001
Defined conditions for DNA extraction from Bacillus subtilis spores journal February 1970
Preparative and analytical purification of DNA from agarose. journal February 1979
A Minisonicator To Rapidly Disrupt Bacterial Spores for DNA Analysis journal October 1999
Radical-generating coordination complexes as tools for rapid and effective fragmentation and fluorescent labeling of nucleic acids for microchip hybridization journal December 2002

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