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Title: Repeat sequence chromosome specific nucleic acid probes and methods of preparing and using

Abstract

A primer directed DNA amplification method to isolate efficiently chromosome-specific repeated DNA wherein degenerate oligonucleotide primers are used is disclosed. The probes produced are a heterogeneous mixture that can be used with blocking DNA as a chromosome-specific staining reagent, and/or the elements of the mixture can be screened for high specificity, size and/or high degree of repetition among other parameters. The degenerate primers are sets of primers that vary in sequence but are substantially complementary to highly repeated nucleic acid sequences, preferably clustered within the template DNA, for example, pericentromeric alpha satellite repeat sequences. The template DNA is preferably chromosome-specific. Exemplary primers and probes are disclosed. The probes of this invention can be used to determine the number of chromosomes of a specific type in metaphase spreads, in germ line and/or somatic cell interphase nuclei, micronuclei and/or in tissue sections. Also provided is a method to select arbitrarily repeat sequence probes that can be screened for chromosome-specificity. 18 figs.

Inventors:
;
Issue Date:
Research Org.:
University of California
OSTI Identifier:
87721
Patent Number(s):
5,427,932
Application Number:
PAN: 7-858,124
Assignee:
Univ. of California, Oakland, CA (United States)
DOE Contract Number:  
W-7405-ENG-48
Resource Type:
Patent
Resource Relation:
Other Information: PBD: 27 Jun 1995
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; DNA SEQUENCERS; DESIGN; GENE AMPLIFICATION; DNA SEQUENCING; OLIGONUCLEOTIDES; MOLECULAR BIOLOGY; DNA

Citation Formats

Weier, H U.G., and Gray, J W. Repeat sequence chromosome specific nucleic acid probes and methods of preparing and using. United States: N. p., 1995. Web.
Weier, H U.G., & Gray, J W. Repeat sequence chromosome specific nucleic acid probes and methods of preparing and using. United States.
Weier, H U.G., and Gray, J W. Tue . "Repeat sequence chromosome specific nucleic acid probes and methods of preparing and using". United States.
@article{osti_87721,
title = {Repeat sequence chromosome specific nucleic acid probes and methods of preparing and using},
author = {Weier, H U.G. and Gray, J W},
abstractNote = {A primer directed DNA amplification method to isolate efficiently chromosome-specific repeated DNA wherein degenerate oligonucleotide primers are used is disclosed. The probes produced are a heterogeneous mixture that can be used with blocking DNA as a chromosome-specific staining reagent, and/or the elements of the mixture can be screened for high specificity, size and/or high degree of repetition among other parameters. The degenerate primers are sets of primers that vary in sequence but are substantially complementary to highly repeated nucleic acid sequences, preferably clustered within the template DNA, for example, pericentromeric alpha satellite repeat sequences. The template DNA is preferably chromosome-specific. Exemplary primers and probes are disclosed. The probes of this invention can be used to determine the number of chromosomes of a specific type in metaphase spreads, in germ line and/or somatic cell interphase nuclei, micronuclei and/or in tissue sections. Also provided is a method to select arbitrarily repeat sequence probes that can be screened for chromosome-specificity. 18 figs.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1995},
month = {6}
}