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US4533628A
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Colony hybridization method
Assignee: New York University
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Filing: 1983-08-03
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Granted: 1985-08-06
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Expires: 2003-08-03
Abstract
Abstract In the colony hybridization technique, bacterial colonies are replica-plated onto filter paper, and their DNA is hybridized with labeled DNA containing a specific sequence. A one-hundred fold increase in sensitivity is obtained by subjecting the colonies to a stream of steam in the presence of alkali for about three minutes prior to hybridization.
Inventors
- 👤 Renata Maas
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 21:40
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6832; C12N; C12N1/00; C12N1/06; C12Q1/6841; Y; Y10; Y10S; Y10S435/00; Y10S435/805; Y10S435/82; Y10T; Y10T436/00; Y10T436/14; Y10T436/142222; Y10T436/143333
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
85 words
A colony hybridization method for detecting a specific DNA sequence comprising the following steps for performing the specified acts: growing bacterial colonies on agar and replicating them onto filter paper, steaming said colonies in the presence of alkali on said paper in order to lyse said colonies on said paper and release their DNA, fixing said DNA to said paper in situ, hybridizing DNA containing said specific sequence to a labelled DNA containing said specific sequence, and detecting said hybridized DNA containing said specific sequence.
Claim 2
Dependent Claim
85 words
A colony hybridization method for detecting a specific DNA sequence comprising the following steps for performing the specified acts: growing bacterial colonies on agar and replicating them onto filter paper, steaming said colonies in the presence of alkali on said paper in order to lyse said colonies on said paper and release their DNA, fixing said DNA to said paper in situ, hybridizing DNA containing said specific sequence to a labelled DNA containing said specific sequence, and detecting said hybridized DNA containing said specific sequence.
Claim 3
Dependent Claim
32 words
The colony hybridization method according to claim 1, wherein said colonies are contacted with a stream of steam in the presence of alkali on said filter paper for at least one minute.
Claim 4
Dependent Claim
32 words
The colony hybridization method according to claim 1, wherein said colonies are contacted with a stream of steam in the presence of alkali for at least two minutes on said filter paper.
Claim 5
Dependent Claim
27 words
The colony hybridization method according to claim 1, wherein said colonies are contacted with a stream of steam in the presence of alkali for about three minutes.
Claim 6
Dependent Claim
16 words
The colony hybridization method according to claim 1, wherein said hybridized DNA is detected by autoradiography.
Claim 7
Dependent Claim
18 words
The colony hybridization method according to claim 5, wherein said hybridized DNA is detected by P 32 -autoradiography.
Claim 8
Dependent Claim
16 words
The colony hybridization method according to claim 1, wherein said hybridized DNA is detected by fluorography.
Claim 9
Dependent Claim
18 words
The colony hybridization method according to claim 7, wherein said hybridized DNA is detected by H 3 -fluorography.
Claim 10
Dependent Claim
27 words
The colony hybridization method according to claim 1, including preparing a reference set of said colonies by replica plating, and using said reference set as a control.
References Cited (Backward Citations)
US4358535A
Cited
US4358535B1
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1988-08-05 | 1989-02-04 | 1989-08-06 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-08-05 | 1993-02-04 | 1993-08-06 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-08-05 | 1997-02-04 | 1997-08-06 | $7700.00 | Window Closed / Lapsed |