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US5447841A
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Methods for chromosome-specific staining
Assignee: University of California Berkeley
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Filing: 1990-12-14
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Granted: 1995-09-05
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Expires: 2010-12-14
Abstract
Abstract Methods and compositions for chromosome-specific staining are provided. Compositions comprise heterogenous mixtures of labeled nucleic acid fragments having substantially complementary base sequences to unique sequence regions of the chromosomal DNA for which their associated staining reagent is specific. Methods include methods for making the chromosome-specific staining compositions of the invention, and methods for applying the staining compositions to chromosomes.
Inventors
- 👤 Joe W. Gray
- 👤 Daniel Pinkel
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 17:13
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6832; C12Q1/6811; C12Q1/6841; C12Q1/6844; C12Q1/6853; C12Q1/6876; C12Q1/6879; C12Q1/6883; C12Q1/6886; C12Q2600/00; C12Q2600/156; Y; Y10; Y10S; Y10S435/00; Y10S435/805; Y10S435/82
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
141 words
A method of staining target chromosomal DNA comprising: (a) providing 1) labeled nucleic acid that comprises fragments which are substantially complementary to nucleic acid segments within the chromosomal DNA for which detection is desired, and 2) blocking nucleic acid that comprises fragments which are substantially complementary to repetitive segments in the labeled nucleic acid; and (b) employing said labeled nucleic acid, blocking nucleic acid, and chromosomal DNA in in situ hybridization so that labeled repetitive segments are substantially blocked from binding to the chromosomal DNA, while hybridization of unique segments within the labeled nucleic acid to the chromosomal DNA is allowed, wherein blocking of the labeled repetitive segments is sufficient to permit detection of hybridized labeled nucleic acid containing unique segments, and wherein the chromosomal DNA is present in a morphologically identifiable chromosome or cell nucleus during the in situ hybridization.
Claim 2
Independent Claim
141 words
A method of staining target chromosomal DNA comprising: (a) providing 1) labeled nucleic acid that comprises fragments which are substantially complementary to nucleic acid segments within the chromosomal DNA for which detection is desired, and 2) blocking nucleic acid that comprises fragments which are substantially complementary to repetitive segments in the labeled nucleic acid; and (b) employing said labeled nucleic acid, blocking nucleic acid, and chromosomal DNA in in situ hybridization so that labeled repetitive segments are substantially blocked from binding to the chromosomal DNA, while hybridization of unique segments within the labeled nucleic acid to the chromosomal DNA is allowed, wherein blocking of the labeled repetitive segments is sufficient to permit detection of hybridized labeled nucleic acid containing unique segments, and wherein the chromosomal DNA is present in a morphologically identifiable chromosome or cell nucleus during the in situ hybridization.
Claim 3
Dependent Claim
29 words
The method of claim 1 wherein the blocking nucleic acid is hybridized with the labeled nucleic acid prior to hybridization of the labeled nucleic acid with the chromosomal DNA.
Claim 4
Dependent Claim
28 words
The method of claim 1 wherein the blocking nucleic acid is hybridized with the chromosomal DNA prior to hybridization of the labeled nucleic acid with the chromosomal DNA.
Claim 5
Dependent Claim
35 words
The method of claim 1 wherein the blocking nucleic acid is hybridized with the labeled nucleic acid and separately with the chromosomal DNA prior to hybridization of the labeled nucleic acid with the chromosomal DNA.
Claim 6
Dependent Claim
26 words
The method of claim 1 wherein the labeled nucleic acid, the blocking nucleic acid, and the chromosomal DNA are hybridized together during in situ hybridization step.
Claim 7
Dependent Claim
33 words
The method of claim 1 wherein the labeled nucleic acid comprises fragments which are designed to allow detection of extra or missing chromosomes, extra or missing portions of a chromosome, or chromosomal rearrangements.
Claim 8
Dependent Claim
15 words
The method of claim 6 wherein the chromosomal rearrangement is a translocation or an inversion.
Claim 9
Dependent Claim
24 words
The method of claim 6 wherein the labeled nucleic acid is designed to allow detection of a chromosomal rearrangement consistent with chronic myelogenous leukemia.
Claim 10
Dependent Claim
17 words
The method of claim 6 wherein the labeled nucleic acid is designed to allow detection of aneuploidy.
Claim 11
Dependent Claim
12 words
The method of claim 6 wherein the extra chromosome is chromosome 21.
Claim 12
Dependent Claim
43 words
The method of claim 1 wherein the labeled nucleic acid comprises fragments complementary to the total genomic complement of chromosomes, fragments complementary to a single chromosome, fragments complementary to a subset of chromosomes, or fragments complementary to a subregion of a single chromosome.
Claim 13
Dependent Claim
24 words
The method of claim 11 wherein the fragments are selected from the nucleic acid of normal human chromosomes 1 through 22, X, and Y.
Claim 14
Dependent Claim
15 words
The method of claim 1 wherein the chromosomal DNA is metaphase or interphase chromosomal DNA.
Claim 15
Dependent Claim
22 words
The method of claim 1 wherein the repetitive segments comprise segments which are substantially complementary to highly repetitive segments and middle-repetitive segments.
Claim 16
Dependent Claim
16 words
The method of claim 14 wherein the repetitive segments are substantially complementary to satellite repetitive segments.
Claim 17
Dependent Claim
25 words
The method of claim 14 wherein the repetitive segments are substantially complementary to tandem repetitive segments or repetitive segments located in regions of centromeric heterochromatin.
Claim 18
Dependent Claim
16 words
The method of claim 14 wherein the repetitive segments are substantially complementary to interspersed repetitive segments.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1998-09-04 | 1999-03-06 | 1999-09-05 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2002-09-04 | 2003-03-06 | 2003-09-05 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2006-09-04 | 2007-03-06 | 2007-09-05 | $7700.00 | Window Closed / Lapsed |