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Apparatus and method of analyzing nucleic acid molecules

Assignee: AUTOSEO Inc
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Filing: 1984-06-08
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Granted: 1989-09-26
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Expires: 2004-06-08
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Abstract

Abstract Apparatus and method of sequencing nucleic acid molecules, including DNA and RNA molecules. Several copies of the molecules are labeled with a radioactive tracer and divided into separate groups. The groups of molecules are then each chemically cleaved to produce labeled fragments of varying length which terminate in predetermined bases. The groups of fragments are then introduced into separate columns of electrophoresis gel and voltage is applied across the gel to cause subgroups of the fragments to propagate along the columns at a velocity which is a function of the length of the fragments in the subgroup. Radiation detectors are used to detect the subgroups of fragments as they pass a predetermined position along the gel column, and data corresponding to the order in which the subgroups of fragments are detected is stored in a memory. Such data is then used to determine the bases sequence of the molecules.

Inventors

  • 👤 Thomas J. Conroy
  • 👤 Martin H. Graham

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-30 12:53
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6869; C12Q1/6813; C12Q1/6816

Claims Analysis

Total Claims: 15
Claim 1
Independent Claim 111 words
A method of analyzing the structure of nucleic acid molecules comprising the following steps: providing several copies of said nucleic acid molecule to be analyzed separating said copies into separate groups of molecule fragments; introducing said groups into separate electrophoresis gel channels; applying a voltage across said gel channels so that each of the groups becomes separating along the channels into subgroups of the fragments, with the fragments which comprise one of the subgroups having the same number of bases. detecting when an individual one of the subgroups passes a predetermined position along said gel channels and producing an output signal in response thereto; and storing data which represent said signals.
Claim 2
Independent Claim 111 words
A method of analyzing the structure of nucleic acid molecules comprising the following steps: providing several copies of said nucleic acid molecule to be analyzed separating said copies into separate groups of molecule fragments; introducing said groups into separate electrophoresis gel channels; applying a voltage across said gel channels so that each of the groups becomes separating along the channels into subgroups of the fragments, with the fragments which comprise one of the subgroups having the same number of bases. detecting when an individual one of the subgroups passes a predetermined position along said gel channels and producing an output signal in response thereto; and storing data which represent said signals.
Claim 3
Dependent Claim 30 words
The method of claim 1 wherein said voltage applied across said gel channels is periodically adjusted to maintain a substantially constant fragment velocity when said subgroups pass said predetermined position.
Claim 4
Dependent Claim 30 words
The method of claim 1 further comprising the step of labeling said molecule fragments with a radioactive tracer and wherein said detecting step is carried out utilizing a radiation detector.
Claim 5
Dependent Claim 84 words
The method of claim 1 wherein said gel channels include a first section where said groups of molecule fragments are introduced followed by a second section at said predetermined position and wherein, subsequent to said step of applying a voltage, said method includes the additional step of increasing the velocity of said subgroups of molecule fragments when said subgroups exit said first section and enter said second section so as to increase the separation between said subgroups of molecule fragments at said predetermined position.
Claim 6
Dependent Claim 47 words
The method of claim 4 wherein said step of increasing the velocity of said subgroups of molecule fragments is carried out by providing said electrophoresis gel channels having said first sections with a relatively large cross-sectional area and said second sections with a relatively small cross-sectional area.
Claim 7
Independent Claim 112 words
A method of analyzing the structure of DNA molecules comprising the following steps: providing several copies of strands of DNA molecules to be analyzed; labeling said copies with a tracer; separating said copies into at least three different groups; chemically cleaving said groups in a predetermined manner into fragments; introducing said fragmented groups into separate electrophoresis gel channels; applying a voltage across said gel channels; detecting said tracer as groups of said fragments having the same number of bases pass a predetermined position along said gel channel; producing a separate output signal for each of said gel channels in response to said detected fragments; and storing data which represent said output signals.
Claim 8
Independent Claim 112 words
A method of analyzing the structure of DNA molecules comprising the following steps: providing several copies of strands of DNA molecules to be analyzed; labeling said copies with a tracer; separating said copies into at least three different groups; chemically cleaving said groups in a predetermined manner into fragments; introducing said fragmented groups into separate electrophoresis gel channels; applying a voltage across said gel channels; detecting said tracer as groups of said fragments having the same number of bases pass a predetermined position along said gel channel; producing a separate output signal for each of said gel channels in response to said detected fragments; and storing data which represent said output signals.
Claim 9
Dependent Claim 24 words
The method of claim 6 wherein said voltage across each of said gel channels is independently controlled and said tracer is a radioactive tracer.
Claim 10
Independent Claim 102 words
A method of analyzing the structure of nucleic acid molecules comprising the following steps: providing fragments of said nucleic acid molecule to be analyzed, the fragments having a varying number of bases; introducing the fragments into an electrophoresis gel channel; applying a voltage across said gel channel so that the fragments become distributed along the channel into separate groups of fragments having the same number of bases; detecting the passage of an individual one of the groups across a predetermined position on said one gel channel and producing an output signal in response thereto; and storing data which represent said output signal.
Claim 11
Independent Claim 102 words
A method of analyzing the structure of nucleic acid molecules comprising the following steps: providing fragments of said nucleic acid molecule to be analyzed, the fragments having a varying number of bases; introducing the fragments into an electrophoresis gel channel; applying a voltage across said gel channel so that the fragments become distributed along the channel into separate groups of fragments having the same number of bases; detecting the passage of an individual one of the groups across a predetermined position on said one gel channel and producing an output signal in response thereto; and storing data which represent said output signal.
Claim 12
Dependent Claim 32 words
The method of claim 8 wherein said voltage applied across said gel channel is periodically adjusted to maintain a substantially constant fragment velocity when said groups of fragments pass said predetermined position.
Claim 13
Dependent Claim 29 words
The method claim 8 further comprising the step of labeling said molecule fragments with a radioactive tracer and wherein said detecting step is carried out utilizing a radiation detector.
Claim 14
Dependent Claim 80 words
The method of claim 8 wherein said gel channel includes a first section where said molecule fragments are introduced followed by a second section at said predetermined position and wherein, subsequent to said step of applying a voltage, said method includes the additional step of increasing the velocity of said molecule fragments when said fragments exit said first section and enter said second section so as to increase the separation between said groups of molecule fragments at said predetermined position.
Claim 15
Dependent Claim 45 words
The method of claim 11 wherein said step of increasing the velocity of said molecule fragments is carried out by providing said electrophoresis gel channel having said first section with a relatively large cross-sectional area and said second section with a relatively small cross-sectional area.

References Cited (Backward Citations)

US4154669A Cited
US4375401A Cited
EP0070959A2 Cited
US4456513A Cited
US4729947A Cited
US4545888A Cited

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1992-09-25 1993-03-27 1993-09-26 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1996-09-25 1997-03-27 1997-09-26 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 2000-09-25 2001-03-27 2001-09-26 $7700.00 Window Closed / Lapsed

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