US4740283A
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Pulsed-field gradient gel electrophoretic apparatus
Assignee: University Patents Inc
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Filing: 1987-02-03
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Granted: 1988-04-26
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Expires: 2007-02-03
Abstract
Abstract A pulsed-field gradient gel electrophoretic apparatus is described wherein a free standing gel is employed. Electrode arrays are oriented to provide three dimensional fields across the face of the gel rather than in the plane of the gel. The electrodes produce three dimensional fields which are orthogonally oriented with respect to each other and cause the molecules to proceed down the lanes in the gel in a generally saw-tooth manner oscillating between the surfaces thereof. Alternately, three dimensional, electric fields are arranged at angles greater than 90° to one another across the face of the gel in separating the molecules, following which transfer electrodes are substituted for the separation electrodes in order to transfer the molecules from their separation lanes onto a separate substrate.
Inventors
- 👤 William A. Laas
- 👤 David Patterson
- 👤 Katheleen J. Gardiner
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 12:17
CPC Classifications
B; B01; B01D; B01D57/00; B01D57/02; G; G01; G01N; G01N27/00; G01N27/26; G01N27/416; G01N27/447; G01N27/44704; G01N27/44713; G01N27/44756; G01N27/44773
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
151 words
Electrophoretic separation apparatus comprising: a medium for supporting particles to be separated, said medium exhibiting at least a pair of opposed generally planar faces separated by a thickness of medium small in relation to the dimension of said faces and including one or more separation track(s); first electrode means positioned about said medium for applying, upon energization, a first electric field across said separation track(s), the direction of said first field intersecting both said planar faces and oriented at an angle which is acute with respect to a first said planar face; second electrode means positioned about said medium for applying, upon energization, a second electric field across said separation track(s), the direction of said second field intersecting both said planar faces and oriented at an angle which is acute with respect to a second said planar face; and means for alternately and repetitively energizing said first and second electrode means.
Claim 2
Dependent Claim
151 words
Electrophoretic separation apparatus comprising: a medium for supporting particles to be separated, said medium exhibiting at least a pair of opposed generally planar faces separated by a thickness of medium small in relation to the dimension of said faces and including one or more separation track(s); first electrode means positioned about said medium for applying, upon energization, a first electric field across said separation track(s), the direction of said first field intersecting both said planar faces and oriented at an angle which is acute with respect to a first said planar face; second electrode means positioned about said medium for applying, upon energization, a second electric field across said separation track(s), the direction of said second field intersecting both said planar faces and oriented at an angle which is acute with respect to a second said planar face; and means for alternately and repetitively energizing said first and second electrode means.
Claim 3
Dependent Claim
22 words
The invention as defined in claim 1 wherein said first and second field directions are approximately orthogonal with respect to each other.
Claim 4
Dependent Claim
52 words
The invention as defined in claim 2, wherein each said first and second electrode means comprises two portions, one portion juxtaposed to one of said pair of opposed generally planar faces of said medium and the other portion to a second of said pair of opposed generally planar faces of said medium.
Claim 5
Dependent Claim
31 words
The invention as defined in claim 3 wherein said one portion includes a conductor grid which produces a three dimensional field configuration which substantially encompasses the length of all separation tracks.
Claim 6
Dependent Claim
30 words
The invention as defined in claim 1 wherein said medium is a gel-like, planar substance through which said particles may move and has sufficient physical rigidity to be free standing.
Claim 7
Dependent Claim
54 words
The invention as defined in claim 5 wherein each said electrode means comprises a multi-conductor, substantially planar array, juxtaposed next to one of said pair of opposed generally planar faces of said medium with a corresponding oppositely poled electrode positioned opposite a second of said pair of opposed generally planar faces of said medium.
Claim 8
Dependent Claim
195 words
Electrophoretic apparatus comprising: a medium for supporting particles to be separated, said medium exhibiting at least a pair of opposed generally planar faces separated by a thickness of medium small in relation to the dimension of said faces and including one or more separation track(s) for advancement of the particles from one end to an opposite end of said separation track(s); first electrode means positioned about said medium for applying, upon energization, a first electric field across said separation track(s), the direction of said first field intersecting both said planar faces adjacent to the one end of said separation track(s) and oriented at an angle which is acute with respect to a first said planar face; second electrode means positioned about said medium for applying, upon energization, a second electric field across said separation track(s), the direction of said second field intersecting both said planar faces adjacent to the one end of said separation track(s) and oriented at an angle which is acute with respect to a second said planar face, the included angle between said electric fields being at least 90°; and means for alternately and repetitively energizing said first and second electrode means.
Claim 9
Dependent Claim
195 words
Electrophoretic apparatus comprising: a medium for supporting particles to be separated, said medium exhibiting at least a pair of opposed generally planar faces separated by a thickness of medium small in relation to the dimension of said faces and including one or more separation track(s) for advancement of the particles from one end to an opposite end of said separation track(s); first electrode means positioned about said medium for applying, upon energization, a first electric field across said separation track(s), the direction of said first field intersecting both said planar faces adjacent to the one end of said separation track(s) and oriented at an angle which is acute with respect to a first said planar face; second electrode means positioned about said medium for applying, upon energization, a second electric field across said separation track(s), the direction of said second field intersecting both said planar faces adjacent to the one end of said separation track(s) and oriented at an angle which is acute with respect to a second said planar face, the included angle between said electric fields being at least 90°; and means for alternately and repetitively energizing said first and second electrode means.
Claim 10
Dependent Claim
31 words
The invention as defined in claim 7 wherein said first and second electrode means have maximum fields at an included angle in the range of 90° to 120° to one another.
Claim 11
Dependent Claim
65 words
The invention as defined in claim 8, wherein said first and second electrode means each comprises a negative electrode disposed adjacent to one planar face of said medium and a positive electrode disposed opposite to the other planar face of said medium, the distance between each said positive electrode and said medium being greater than the distance between each said negative electrode and said medium.
Claim 12
Dependent Claim
64 words
The invention as defined in claim 7 wherein said medium is an upright, gel-like, planar substance centered between said first and second electrode means, said medium having wells in communication with the one end of said separation track(s), said first and second electric fields extending in a downward angular direction through the thickness of said medium between the opposite ends of said separation track(s).
Claim 13
Dependent Claim
29 words
The invention as defined in claim 10, said first and second electrode means each defined by at least one wire extending horizontally in spaced parallel relation to said medium.
Claim 14
Dependent Claim
69 words
The invention as defined in claim 10, said apparatus including a common housing having medium support means for removably supporting said planar medium symmetrically between said negative and positive electrodes, electrode support means for removably supporting said negative and positive electrodes in said housing, and transfer electrode means including means removably positioning said transfer electrode means in said housing for applying upon energization a uni-directional field across said medium.
Claim 15
Dependent Claim
32 words
The invention as defined in claim 12, said transfer electrode means operative to apply a progressively reduced voltage drop across said medium from one end to the opposite end of said medium.
Claim 16
Dependent Claim
35 words
The invention as defined in claim 13, said transfer electrode means including positive and negative electrodes each in the form of a multi-conductor array diverging downwardly and away from opposite planar faces of said medium.
Claim 17
Independent Claim
115 words
The method of performing separation and transfer of macromolecular particles from separation track(s) of a supporting medium onto a substrate comprising the steps of: (1) orienting said medium in a vertical plane symmetrically between intersecting first and second electrical fields; (2) alternately and repetitively directing said first and second electrical fields at downwardly directed angles through the upper end of said supporting medium until said macromolecular particles are separated along said separation track(s); (3) applying a substrate to one planar surface of said medium; and (4) exposing said medium to a third electric field extending across said medium in a direction causing said macromolecular particles to be transferred from said separation track(s) onto said substrate.
Claim 18
Independent Claim
115 words
The method of performing separation and transfer of macromolecular particles from separation track(s) of a supporting medium onto a substrate comprising the steps of: (1) orienting said medium in a vertical plane symmetrically between intersecting first and second electrical fields; (2) alternately and repetitively directing said first and second electrical fields at downwardly directed angles through the upper end of said supporting medium until said macromolecular particles are separated along said separation track(s); (3) applying a substrate to one planar surface of said medium; and (4) exposing said medium to a third electric field extending across said medium in a direction causing said macromolecular particles to be transferred from said separation track(s) onto said substrate.
Claim 19
Dependent Claim
22 words
The method according to claim 15, in which the included angle between said first and second electric fields is at least 90°.
Claim 20
Dependent Claim
29 words
The method according to claim 15, characterized in step (4) by applying a progressively reduced voltage drop across said medium from the upper end to the lower end thereof.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-04-26 | 1991-10-26 | 1992-04-26 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-04-26 | 1995-10-26 | 1996-04-26 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-04-26 | 1999-10-26 | 2000-04-26 | $7700.00 | Window Closed / Lapsed |