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Title: Method for isolating chromosomal DNA in preparation for hybridization in suspension

Abstract

A method is provided for detecting nucleic acid sequence aberrations using two immobilization steps. According to the method, a nucleic acid sequence aberration is detected by detecting nucleic acid sequences having both a first nucleic acid sequence type (e.g., from a first chromosome) and a second nucleic acid sequence type (e.g., from a second chromosome), the presence of the first and the second nucleic acid sequence type on the same nucleic acid sequence indicating the presence of a nucleic acid sequence aberration. In the method, immobilization of a first hybridization probe is used to isolate a first set of nucleic acids in the sample which contain the first nucleic acid sequence type. Immobilization of a second hybridization probe is then used to isolate a second set of nucleic acids from within the first set of nucleic acids which contain the second nucleic acid sequence type. The second set of nucleic acids are then detected, their presence indicating the presence of a nucleic acid sequence aberration. Chromosomal DNA in a sample containing cell debris is prepared for hybridization in suspension by treating the mixture with RNase. The treated DNA can also be fixed prior to hybridization.

Inventors:
 [1]
  1. San Ramon, CA
Issue Date:
Research Org.:
Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)
OSTI Identifier:
873044
Patent Number(s):
6077671
Assignee:
Regents of University of California (Oakland, CA)
Patent Classifications (CPCs):
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
DOE Contract Number:  
W-7405-ENG-48
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
method; isolating; chromosomal; dna; preparation; hybridization; suspension; provided; detecting; nucleic; acid; sequence; aberrations; immobilization; steps; according; aberration; detected; sequences; type; chromosome; presence; indicating; probe; isolate; set; acids; sample; contain; containing; cell; debris; prepared; treating; mixture; rnase; treated; fixed; prior; sequence aberration; detecting nucleic; sequence aberrations; sequence type; sample containing; chromosomal dna; acid sequence; nucleic acid; acid sequences; nucleic acids; hybridization probe; sequence indicating; immobilization steps; /435/999/

Citation Formats

Lucas, Joe N. Method for isolating chromosomal DNA in preparation for hybridization in suspension. United States: N. p., 2000. Web.
Lucas, Joe N. Method for isolating chromosomal DNA in preparation for hybridization in suspension. United States.
Lucas, Joe N. Sat . "Method for isolating chromosomal DNA in preparation for hybridization in suspension". United States. https://www.osti.gov/servlets/purl/873044.
@article{osti_873044,
title = {Method for isolating chromosomal DNA in preparation for hybridization in suspension},
author = {Lucas, Joe N},
abstractNote = {A method is provided for detecting nucleic acid sequence aberrations using two immobilization steps. According to the method, a nucleic acid sequence aberration is detected by detecting nucleic acid sequences having both a first nucleic acid sequence type (e.g., from a first chromosome) and a second nucleic acid sequence type (e.g., from a second chromosome), the presence of the first and the second nucleic acid sequence type on the same nucleic acid sequence indicating the presence of a nucleic acid sequence aberration. In the method, immobilization of a first hybridization probe is used to isolate a first set of nucleic acids in the sample which contain the first nucleic acid sequence type. Immobilization of a second hybridization probe is then used to isolate a second set of nucleic acids from within the first set of nucleic acids which contain the second nucleic acid sequence type. The second set of nucleic acids are then detected, their presence indicating the presence of a nucleic acid sequence aberration. Chromosomal DNA in a sample containing cell debris is prepared for hybridization in suspension by treating the mixture with RNase. The treated DNA can also be fixed prior to hybridization.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Sat Jan 01 00:00:00 EST 2000},
month = {Sat Jan 01 00:00:00 EST 2000}
}

Works referenced in this record:

A chromosome painting method for human sperm chromosomes using fluorescentin situ hybridization
journal, June 1994


DNA-binding proteins on lampbrush chromosome loops
journal, May 1989