Nucleic acid purification system and method

• US 5,217,593 A
• Filed: 07/10/1992
• Issued: 06/08/1993
• Est. Priority Date: 03/13/1991
• Status: Expired due to Term

• Current Assignee: MacConnell, William P.
• Original Assignee: MacConnell, William P.
• Inventors: MacConnell, William P.
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US07/911,515
• Time in Patent Office: 333 Days
• Field of Search: 204/182.8, 204/301, 204/299 R, 204/182.1, 204/182.9
• US Class Current: 204/457
• CPC Class Codes: G01N 27/4473 by electric means
• CPC Class Codes: G01N 27/44756 Apparatus specially adapted therefor

Abstract

A process for the purification of DNA and the like comprises a housing having walls forming a reservoir having a chamber for containing a buffer solution, means for circulating a buffer through the reservoir, a disposable cassette within said chamber having first means including a gel for defining a first path extending between an inlet end and an outlet end, a well for introducing a bacterial sample into the path at said inlet end thereof, and a second path intersecting the first path via an elution chamber, having a collection chamber including an elution window at said outlet end, and an electrical circuit for selectively applying an electrical potential along each of the paths for selectively moving a plasmid first along the first path from the bacterial well to the elution chamber, then along the second path to the collection window at the end thereof.

Claims

1. A purification apparatus for the purification of DNA and the like comprising:

a housing forming a chamber having a front end and a back end for containing a buffer solution;

means for circulating a buffer within said chamber;

disposable cassette means positionable within said housing chamber having a front end and a back end having first means including a gel for defining a first path extending between said front end and said back end, inlet means at said front end for introducing a bacterial sample into said path at said front end thereof, an outlet means communicating with said gel at said back end, and second means for defining a second path for extending from said outlet means of said first path via an elution chamber at said back end to a collection chamber, said disposable cassette means comprises a first body formed of a non-conductive material with spaced apart upper and lower panels for containing said gel for defining said first path, said inlet means for introducing a sample at said front end comprises an opening in said upper panel, said second means comprises a second body having a passage defining said elution chamber and having an inlet for communicating with said outlet, and selectively moveable into and out of a position in communication with said outlet means; and

means for selectively applying an electrical potential along each of said paths for selectively moving a plasmid first along said first path from said bacterial sample to the intersection of said first path then along said second path to said collection window at the end thereof.

2. A purification apparatus according to claim 1 wherein said inlet means includes means extending upward from around said opening for defining said inlet means, upstanding walls surrounding said outlet for defining a well for receiving said second body.

3. A purification apparatus according to claim 2 wherein said first body is formed of a plurality of said first paths and said second body is formed with a plurality of said second paths.

4. A purification apparatus according to claim 3 wherein said first body has a generally rectangular box-like configuration and said gel is an electrophoretic agarose gel.

19. A purification apparatus according to claim 1 wherein said means for selectively applying an electrical potential along each of said paths comprises a source of variable electrical power, and means for selectively varying said electrical power.

20. A purification apparatus according to claim 19 wherein said control means includes a programmable CPU and means for programming said CPU for periodic adjustment of said electrical potential.

5. A purification apparatus for the purification of DNA and the like comprising:

a housing forming a chamber having a front end and a back end for containing a buffer solution;

means for circulating a buffer within said chamber;

disposable cassette means positionable within said housing chamber having a front end and a back end and having first means including a gel for defining a first path extending between said front end and said back end, inlet means at said front end for introducing a bacterial sample into said path at said front end thereof, an outlet means communicating with said gel at said back end, and second means for defining a second path for extending from said outlet means of said first path via an elution chamber at said back end to a collection chamber, said first means is a body having a generally rectangular box-like configuration and is formed of a plurality of said first paths and said second means is a body having a plurality of said second paths; and

means for selectively applying an electrical potential along each of said paths for selectively moving a plasmid first along said first path from said bacterial sample to the intersection of said first path then along said second path to said collection window at the end thereof.

6. A purification apparatus according to claim 5 wherein said means for selectively applying an electrical potential along each of said paths comprises a source of variable electrical power, and means for selectively varying said electrical power.

21. A purification apparatus according to claim 5 wherein said inlet means includes means extending upward from around said opening for defining said inlet means, upstanding walls surrounding said outlet for defining a well for receiving said second body.

22. A purification apparatus according to claim 6 wherein said control means includes a programmable CPU and means for programming said CPU for periodic adjustment of said electrical potential.

7. A purification apparatus for the purification of DNA and the like comprising:

a housing forming a chamber having a front end and a back end for containing a buffer solution;

means for circulating a buffer within said chamber;

disposable cassette means positionable within said housing chamber having a front end and a back end having first means including a gel for defining a first path extending between said front end and said back end, inlet means at said front end for introducing a bacterial sample into said path at said front end thereof, an outlet means communicating with said gel at said back end, and second means for defining a second path for extending from said outlet means of said first path via an elution chamber at said back end to a collection chamber, said disposable cassette means comprises a first body formed of a non-conductive material with spaced apart upper and lower panels, a plurality of separating walls between said upper and lower panels defining channels for containing said gel and defining a plurality of said first paths, said inlet means for introducing a sample at said front end comprises an opening in said upper panel into each of said channels, said second means comprises a second body having a plurality of passages defining a plurality of said elution chambers, each having an inlet for communicating with an outlet of said channels, and said second body selectively moveable into a position in communication with said outlet means; and

means for selectively applying an electrical potential along each of said paths for selectively moving a plasmid first along said first path from said bacterial sample to the intersection of said first path then along said second path to said collection window at the end thereof.

8. A purification apparatus according to claim 7 wherein said means for selectively applying an electrical potential along each of said paths comprises a source of variable electrical power, and means for selectively varying said electrical power.

9. A purification apparatus according to claim 8 wherein said control means includes a programmable CPU and means for programming said CPU for periodic adjustment of said electrical potential.

10. An apparatus for the purification of DNA and the like comprising:

a generally box-like housing having a chamber for containing a buffer solution;

means for circulating a buffer to said chamber;

a disposable cassette detachably mountable within said chamber and having a pair of vertically spaced horizontally extending walls, an electrophoretic gel disposed between said walls for defining at least a part of a first path extending generally horizontally between an inlet at a front end and an outlet at a back end of said chamber, an inlet well for introducing a bacterial sample into said path at said inlet end thereof, and an elution well at said outlet;

an elution chamber assembly comprising a body having a passage defining an elution chamber and a collection chamber positionable in said elution well in communication with said outlet for defining a second path intersecting said first path and extending to said collection chamber; and

means for selectively applying an electrical potential along each of said paths for selectively moving a plasmid first along said first path from said inlet well to said elution chamber, then along said second path to said collection chamber at the end thereof.

11. A purification apparatus according to claim 10 wherein said disposable cassette includes a plurality of vertically extending walls disposed between said vertically spaced horizontally extending walls forming a plurality of said first paths, said body includes passage means forming a plurality of said second paths and a collection window comprising a dialysis membrane covering an outlet opening to said passage means.

12. A purification apparatus according to claim 11 wherein said plurality of vertically extending walls extend beyond said outlet into said elution well;

and

said body includes a plurality of slots in a wall thereof for receiving the walls extending into said elution well.

13. A purification apparatus according to claim 12 wherein said means for selectively applying an electrical potential along each of said paths comprises a source of variable electrical power, and means for selectively varying said electrical power.

14. A purification apparatus according to claim 11 wherein said control means includes a programmable CPU and means for programming said CPU for periodic adjustment of said electrical potential.

15. A process for the purification of DNA and the like comprising the steps of:

providing a housing having walls forming a chamber having a front and a back for containing a buffer solution;

providing disposable cassette means having an inlet end with inlet means and an outlet end with outlet means and a gel defining a first path extending from said inlet to said outlet, and elution chamber means having an elution chamber and a collection chamber moveable to a position for defining a second path intersecting said first path at said outlet;

introducing a bacterial sample into said path at said inlet means;

circulating a buffer into said chamber in contact with said gel; and

selectively applying an electrical potential along each of said paths for selectively moving a plasmid along said first path from said bacterial sample to said outlet, moving said elution means into position intersecting said first path, applying an electrical potential along said second path for moving said plasmid along said second path to said collection window at the end thereof.

16. A purification process according to claim 15 wherein:

said disposable cassette means comprises a first body formed of a non-conductive material with spaced apart upper and lower panels, a plurality of separating walls between said upper and lower panels defining channels for containing said gel and defining a plurality of said first paths, said inlet means for introducing a sample at said front end comprises an opening in said upper panel into each of said channels; and

said elution chamber means comprises a second body having a plurality of passages defining a plurality of said elution chambers, each having an inlet for communicating with an outlet of said channels.

17. A purification process according to claim 16 wherein said applying an electrical potential along each of said paths is carried out by providing a source of variable electrical power, and control means for selectively varying said electrical power.

18. A purification process according to claim 17 wherein said control means includes a programmable CPU and means for programming said CPU for periodic adjustment of said electrical potential.