Reaction chamber

• US 8,999,636 B2
• Filed: 01/08/2008
• Issued: 04/07/2015
• Est. Priority Date: 01/08/2007
• Status: Expired due to Fees

• Current Assignee: Toxic Reports LLC
• Original Assignee: Toxic Reports LLC
• Inventors: Patil, Vishal, Mollova, Emilia, Gilmanshin, Rudolf
• Primary Examiner(s): FORMAN, BETTY J
• Application Number:
  US12/448,800
• Publication Number:
  US 20100120101A1
• Time in Patent Office: 2,646 Days
• Field of Search: None
• US Class Current: 435/6.1
• CPC Class Codes: C12N 15/1003 Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor

Abstract

A chamber in which an agent, like genomic DNA, may be harvested and optionally manipulated rapidly (e.g., on the order of a few hours), without shearing or fragmentation.

Claims

1. A system for separating agents from non-agents in a chamber, the system comprising:

a chamber comprising;

a fluid introduction port;

a porous substrate positioned substantially opposite from the fluid introduction port in the chamber, the porous substrate configured to prevent the passage of agents that are larger than a threshold size of the porous substrate, the porous substrate having a first side and a second side, wherein the fluid introduction port is positioned on the first side of the porous substrate;

a plurality of ports coupled to the chamber and configured to permit fluid to exit the chamber, wherein the plurality of ports includes at least a first port, a second port, and a third port, wherein the first port and the second port are positioned on the first side of the porous substrate and the third port is positioned on the second side of the porous substrate, and wherein the first port is substantially parallel to a surface of the porous substrate;

a controller configured to control;

a first flow of fluid into the chamber from the fluid introduction port and through the porous substrate to hold agents and other constituents at the porous substrate; and

a second flow of fluid into the chamber from the fluid introduction port and through at least one of the first and second ports to allow other constituents to move away from the agents on the porous substrate.

2. The system of claim 1, wherein the controller is configured to reduce the second flow relative to the first flow to direct agents towards the porous substrate when agents are introduced to the chamber through the fluid introduction port.

3. The system of claim 1, wherein the controller is configured to increase the second flow relative to the first flow to move other constituents away from the agents on the porous substrate.

4. The system of claim 1, wherein the controller is configured to substantially stop the first and second flows through the chamber to allow reactions to occur between the agents and other constituents.

5. The system of claim 1, wherein the controller is configured to reverse the first flow to move the agents away from the porous substrate and towards the fluid introduction port.

6. The system of claim 1, wherein the agents comprise nucleic acids that have been separated from one or more cells from which the nucleic acids were originally presented.

7. The system of claim 1, wherein the other constituents are selected from the group comprising:

sequence specific probes, intercalators, cellular debris, lytic enzymes, endonucleases, and exonucleases.

8. The system of claim 1, wherein the agents comprise proteins.

9. The system of claim 1, wherein the controller is configured to automatically control the first flow and the second flow according to an operating protocol.

10. The system of claim 1, wherein the chamber has a substantially truncated cone shape.

11. A system for separating agents from non-agents in a chamber, the system comprising:

a chamber comprising;

a fluid introduction port;

a porous substrate positioned substantially opposite from the fluid introduction port in the chamber, the porous substrate configured to prevent the passage of agents that are larger than a threshold size of the porous substrate, the porous substrate having a first side and a second side, wherein the fluid introduction port is positioned on the first side of the porous substrate;

a plurality of ports coupled to the chamber and configured to permit fluid to exit the chamber, wherein the plurality of ports includes at least a first port, a second port, a third port and a fourth port, wherein the first port and the second port are positioned on the first side of the porous substrate and the third port and the fourth port are positioned on the second side of the porous substrate;

a controller configured to control;

a first flow of fluid into the chamber from the fluid introduction port and through the porous substrate to hold agents and other constituents at the porous substrate; and

a second flow of fluid into the chamber from the fluid introduction port and through at least one of the first and second ports to allow other constituents to move away from the agents on the porous substrate.

12. The system of claim 11, wherein the chamber has a substantially truncated cone shape.

13. A system for separating agents from non-agents in a chamber, the system comprising:

a chamber with a substantially truncated cone shape, the chamber comprising;

a fluid introduction port;

a porous substrate positioned substantially opposite from the fluid introduction port in the chamber, the porous substrate configured to prevent the passage of agents that are larger than a threshold size of the porous substrate, the porous substrate having a first side and a second side, wherein the fluid introduction port is positioned on the first side of the porous substrate;

a plurality of ports coupled to the chamber and configured to permit fluid to exit the chamber, wherein the plurality of ports includes at least a first port, a second port and a third port, wherein the first port and the second port are positioned on the first side of the porous substrate and the third port is positioned on the second side of the porous substrate;

a controller configured to control;

a first flow of fluid into the chamber from the fluid introduction port and through the porous substrate to hold agents and other constituents at the porous substrate; and

a second flow of fluid into the chamber from the fluid introduction port and through at least one of the first and second ports to allow other constituents to move away from the agents on the porous substrate,

wherein the controller is configured to reduce the second flow relative to the first flow to direct agents towards the porous substrate when agents are introduced to the chamber through the fluid introduction port.

14. A system for separating agents from non-agents in a chamber, the system comprising:

a chamber with a substantially truncated cone shape, the chamber comprising;

a fluid introduction port;

a porous substrate positioned substantially opposite from the fluid introduction port in the chamber, the porous substrate configured to prevent the passage of agents that are larger than a threshold size of the porous substrate, the porous substrate having a first side and a second side, wherein the fluid introduction port is positioned on the first side of the porous substrate;

a plurality of ports coupled to the chamber and configured to permit fluid to exit the chamber, wherein the plurality of ports includes at least a first port, a second port and a third port, wherein the first port and the second port are positioned on the first side of the porous substrate and the third port is positioned on the second side of the porous substrate;

a controller configured to control;

a first flow of fluid into the chamber from the fluid introduction port and through the porous substrate to hold agents and other constituents at the porous substrate; and

a second flow of fluid into the chamber from the fluid introduction port and through at least one of the first and second ports to allow other constituents to move away from the agents on the porous substrate,

wherein the controller is configured to increase the second flow relative to the first flow to move other constituents away from the agents on the porous substrate.