Repeat sequence chromosome specific nucleic acid probes and methods of preparing and using
- Tracy, CA
- San Francisco, CA
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.
- Research Organization:
- Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)
- DOE Contract Number:
- W-7405-ENG-48
- Assignee:
- Reagents of University of California (Oakland, CA)
- Patent Number(s):
- US 5427932
- OSTI ID:
- 869950
- Country of Publication:
- United States
- Language:
- English
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sequence
chromosome
specific
nucleic
acid
probes
methods
preparing
primer
directed
dna
amplification
method
isolate
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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
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acid probes
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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
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