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 ard 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.

Inventors:

Weier, Heinz-Ulrich G ^[1]; Gray, Joe W ^[2]

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+ Show Inventor Affiliations

1. Tracy, CA
2. San Francisco, CA

Issue Date:

Sun Jan 01 00:00:00 EST 1995

Research Org.:

Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)

OSTI Identifier:

869950

Patent Number(s):

5427932

Assignee:

Reagents of University of California (Oakland, CA)

Patent Classifications (CPCs):

C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS

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

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C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS

C12Q1/6841 - In situ hybridisation
C12Q1/686 - Polymerase chain reaction [PCR]
C12Q1/6883 - for diseases caused by alterations of genetic material
C12Q2525/151 - repeat or repeated sequences, e.g. VNTR, microsatellite, concatemer
C12Q2525/179 - incorporating arbitrary or random nucleotide sequences

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

Y10S435/81 - Packaged device or kit

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DOE Contract Number:  

W-7405-ENG-48

Resource Type:

Patent

Country of Publication:

United States

Language:

English

Subject:

repeat; sequence; chromosome; specific; nucleic; acid; probes; methods; preparing; primer; directed; dna; amplification; method; isolate; efficiently; chromosome-specific; repeated; degenerate; oligonucleotide; primers; disclosed; produced; heterogeneous; mixture; blocking; staining; reagent; elements; screened; specificity; size; degree; repetition; parameters; sets; vary; substantially; complementary; highly; sequences; preferably; clustered; template; example; pericentromeric; alpha; satellite; exemplary; determine; chromosomes; type; metaphase; spreads; germ; line; somatic; cell; interphase; nuclei; micronuclei; tissue; sections; provided; select; arbitrarily; chromosome-specificity; acid probes; acid sequence; nucleic acid; acid sequences; chromosome specific; acid probe; chromosome-specific staining; specific nucleic; repeat sequences; repeat sequence; amplification method; germ line; substantially complementary; staining reagent; specific type; oligonucleotide primers; interphase nuclei; oligonucleotide primer; dna amplification; /435/436/999/