US4900416A
Expired / Lapsed
US
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Electrophoretic method
Assignee: Fujifilm Holdings Corp
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Filing: 1988-09-13
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Granted: 1990-02-13
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Expires: 2008-09-13
Abstract
Abstract An electrophoretic method comprises the steps of positioning a sample, which contains nucleic acid fragments having different numbers of bases, in an electrophoresis medium composed of an electrophoresis gel medium containing an aqueous poly(meth)acrylamide gel prepared by cross-linking polymerization of a (meth)acrylamide compound and a cross-linking agent in the presence of water and a compound having at least one carbamoyl group as a denaturing agent (or modifier), and applying pulsed electric fields to the electrophoresis medium in two directions, thereby to move the nucleic acid fragments in the presence of molecular sieve effects and to separate the nucleic acid fragments.
Inventors
- 👤 Yoshihiko Makino
- 👤 Masashi Ogawa
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 16:46
CPC Classifications
G; G01; G01N; G01N27/00; G01N27/26; G01N27/416; G01N27/447; G01N27/44704; G01N27/44747
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
131 words
An electrophoretic method comprising the steps of: (i) positioning a sample, which contains nucleic acid fragments having different numbers of bases, in an electrophoresis medium composed of an electrophoresis gel medium containing an aqueous poly(meth)acrylamide gel prepared by the cross-linking polymerization of a (meth)acrylamide compound and a cross-linking agent in the presence of water and a compound having at least one carbamoyl group as a denaturing agent, and (ii) applying pulsed electric fields to said electrophoresis medium in two directions, the electric field strength in each direction being from about 10 V/cm to 150 V/cm, and the duration of each pulse being from about 0.1 msec to 100 msec, to move thereby said nucleic acid fragments in the presence of a molecular sieve effect and to separate said nucleic acid fragments.
Claim 2
Dependent Claim
131 words
An electrophoretic method comprising the steps of: (i) positioning a sample, which contains nucleic acid fragments having different numbers of bases, in an electrophoresis medium composed of an electrophoresis gel medium containing an aqueous poly(meth)acrylamide gel prepared by the cross-linking polymerization of a (meth)acrylamide compound and a cross-linking agent in the presence of water and a compound having at least one carbamoyl group as a denaturing agent, and (ii) applying pulsed electric fields to said electrophoresis medium in two directions, the electric field strength in each direction being from about 10 V/cm to 150 V/cm, and the duration of each pulse being from about 0.1 msec to 100 msec, to move thereby said nucleic acid fragments in the presence of a molecular sieve effect and to separate said nucleic acid fragments.
Claim 3
Dependent Claim
27 words
A method as defined in claim 1 wherein said pulsed electric fields in two directions are opposite to each other and do not substantially intersect each other.
Claim 4
Dependent Claim
22 words
A method as defined in claim 1 wherein said pulsed electric fields in two directions are alternately applied with a predetermined period.
Claim 5
Dependent Claim
33 words
A method as defined in claim 3 wherein the pulsed electric field applied in the direction that causes said nucleic acid fragments to move forward is applied for a relatively longer duration time.
Claim 6
Dependent Claim
22 words
A method as defined in claim 1 wherein said pulsed electric fields in two directions are alternately applied with substantially equal strengths.
Claim 7
Dependent Claim
32 words
A method as defined in claim 1 wherein said electrophoresis medium is a thin plate-shaped or thin membrane-shaped electrophoresis medium membrane having a substantially constant gel concentration and a substantially constant thickness.
Claim 8
Dependent Claim
47 words
A method as defined in claim 1 wherein said electrophoresis medium is a thin plate-shaped or thin membrane-shaped electrophoresis medium membrane provided with at least one gradient selected from the group consisting of a membrane thickness gradient, a gel concentration gradient and a pH buffer concentration gradient.
Claim 9
Independent Claim
26 words
A method as defined in any of claims 1 to 7 wherein said electrophoresis medium is provided on a sheet-shaped support formed of an organic polymer.
Claim 10
Independent Claim
24 words
A method as defined in any of claims 1 to 7 wherein said electrophoresis medium is provided on a sheet-shaped support formed of glass.
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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) | 1993-02-12 | 1993-08-14 | 1994-02-13 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-02-12 | 1997-08-14 | 1998-02-13 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-02-12 | 2001-08-14 | 2002-02-13 | $7700.00 | Window Closed / Lapsed |