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

Patent ·
OSTI ID:873044

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.

Research Organization:
Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)
DOE Contract Number:
W-7405-ENG-48
Assignee:
Regents of University of California (Oakland, CA)
Patent Number(s):
US 6077671
OSTI ID:
873044
Country of Publication:
United States
Language:
English

References (3)

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
Detection of chromosome aberrations in metaphase and interphase tumor cells by in situ hybridization using chromosome-specific library probes journal November 1988