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

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:
 [1];  [2]
  1. (Tracy, CA)
  2. (San Francisco, CA)
Issue Date:
OSTI Identifier:
869950
Assignee:
Reagents of University of California (Oakland, CA) LLNL
Patent Number(s):
US 5427932
Contract Number:
W-7405-ENG-48
Research Org:
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
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/