Nucleic acid isolation process

Longmire, Jonathan L; Lewis, Annette K; Hildebrand, Carl E

Title: Nucleic acid isolation process

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Abstract

A method is provided for isolating DNA from eukaryotic cell and flow sorted chromosomes. When DNA is removed from chromosome and cell structure, detergent and proteolytic digestion products remain with the DNA. These products can be removed with organic extraction, but the process steps associated with organic extraction reduce the size of DNA fragments available for experimental use. The present process removes the waste products by dialyzing a solution containing the DNA against a solution containing polyethylene glycol (PEG). The waste products dialyze into the PEG leaving isolated DNA. The remaining DNA has been prepared with fragments containing more than 160 kb. The isolated DNA has been used in conventional protocols without affect on the protocol.

Inventors:

Longmire, Jonathan L ^[1]; Lewis, Annette K ^[2]; Hildebrand, Carl E ^[1]

Los Alamos, NM
La Jolla, CA

Issue Date:: 1990-01-01

Research Org.:: Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

OSTI Identifier:: 867364

Patent Number(s):: 4921952

Assignee:: United States of America as represented by United States (Washington, DC)

Patent Classifications (CPCs):: 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

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

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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
Y10S435/803 - Physical recovery methods, e.g. chromatography, grinding

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
C12N15/1003 - {Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods

C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
C12Q1/6806 - Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay

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DOE Contract Number:: W-7405-ENG-36

Resource Type:: Patent

Country of Publication:: United States

Language:: English

Subject:: nucleic; acid; isolation; process; method; provided; isolating; dna; eukaryotic; cell; flow; sorted; chromosomes; removed; chromosome; structure; detergent; proteolytic; digestion; products; remain; organic; extraction; steps; associated; reduce; size; fragments; available; experimental; removes; waste; dialyzing; solution; containing; polyethylene; glycol; peg; dialyze; leaving; isolated; remaining; prepared; 160; kb; conventional; protocols; affect; protocol; waste product; cell structure; ethylene glycol; polyethylene glycol; isolated dna; solution containing; nucleic acid; dna fragments; waste products; dna fragment; organic extraction; containing poly; process removes; process step; flow sorted; process steps; acid isolation; /999/435/

Citation Formats


                    Longmire, Jonathan L, Lewis, Annette K, and Hildebrand, Carl E. Nucleic acid isolation process.  United States: N. p., 1990. 
        Web.

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                    Longmire, Jonathan L, Lewis, Annette K, & Hildebrand, Carl E. Nucleic acid isolation process.  United States.

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                    Longmire, Jonathan L, Lewis, Annette K, and Hildebrand, Carl E. Mon .  
        "Nucleic acid isolation process".  United States.  https://www.osti.gov/servlets/purl/867364.

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@article{osti_867364,

  title        = {Nucleic acid isolation process},

  author       = {Longmire, Jonathan L and Lewis, Annette K and Hildebrand, Carl E},

  abstractNote = {A method is provided for isolating DNA from eukaryotic cell and flow sorted chromosomes. When DNA is removed from chromosome and cell structure, detergent and proteolytic digestion products remain with the DNA. These products can be removed with organic extraction, but the process steps associated with organic extraction reduce the size of DNA fragments available for experimental use. The present process removes the waste products by dialyzing a solution containing the DNA against a solution containing polyethylene glycol (PEG). The waste products dialyze into the PEG leaving isolated DNA. The remaining DNA has been prepared with fragments containing more than 160 kb. The isolated DNA has been used in conventional protocols without affect on the protocol.},

  doi          = {},

  journal      = {},
number       = ,

  volume       = ,

  place        = {United States},

  year         = {1990},

  month        = {1}

}

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