FOCUSING CHAMBER

• US 20100112576A1
• Filed: 10/05/2009
• Published: 05/06/2010
• Est. Priority Date: 10/03/2008
• Status: Active Grant

• Current Assignee: PathoGenetix, Inc. (Toxic Reports LLC)
• Original Assignee: U.S. Genomics, Inc (Toxic Reports LLC)
• Inventors: Patil, Vishal
• Granted Patent: US 8,361,716 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: G01N 1/40 Concentrating samples
• CPC Class Codes: G01N 2001/4088 filtration

Abstract

Aspects of the invention relate to devices and methods of use thereof for concentrating, positioning and/or manipulating agents within a fluid, including but not limited to genomic DNA.

Claims

1. A method positioning agents in a fluid flow, the method comprising:

providing a chamber having a substrate;

flowing a first fluid containing agents through a first fluid port and through the substrate in the chamber;

flowing a second fluid flow through one or more second fluid ports positioned about the substrate in the chamber, the second fluid flow at least partially surrounding the first fluid flow to direct the first fluid flow and any agents contained therein toward a central portion of the substrate;

positioning the agents on the central portion of the substrate;

reversing flow through the first fluid port to move any agents positioned on the substrate out of the chamber in central streamlines that exit the chamber through the first fluid port; and

removing flow from about the central streamlines to separate fluid flow that lacks agents from the central streamlines that exit the chamber.

2. The method of claim 1, wherein flowing the second fluid comprises flowing the second fluid through one or more second fluid ports positioned about the substrate in the chamber, the second fluid flow completely surrounding the first fluid flow to direct the first fluid flow and any agents contained therein toward the central portion of the substrate.

3. The method of claim 1, wherein removing flow from about the central streamlines to separate fluid flow that lacks agents from the central streamlines that exit the chamber comprises removing flow through the one or more second fluid ports.

4. The method of claim 1, wherein removing flow from about the central streamlines to separate fluid flow that lacks agents from the central streamlines that exit the chamber comprises removing flow through the one or more third fluid ports that are positioned between the first fluid port and the one or more second fluid ports.

5. The method of claim 4, wherein flow through the second ports is prevented when reversing flow through the first fluid port.

6. The method of claim 4, wherein flow is introduced through the second ports when reversing flow through the first fluid port.

7. The method of claim 1, wherein providing the chamber comprises providing a chamber that includes an open top.

8. The method of claim 1, wherein a flow rate of second fluid is five times or greater than a flow rate of the second fluid.

9. The method of claim 1, wherein a flow rate of the second fluid is fifteen times or greater than a flow rate of the second fluid.

10. The method of claim 1, wherein a flow rate of the second fluid is twenty times or greater than a flow rate of the second fluid.

11. The method of claim 1, wherein agents removed from the chamber are concentrated ten times greater than prior to entering the chamber.

12. The method of claim 1, wherein agents removed from the chamber are concentrated one hundred times greater than prior to entering the chamber.

13. The method of claim 1, wherein the agents are nucleic acids.

14. The method of claim 13, wherein the agents are DNA.

15. The method of claim 13, wherein the nucleic acids maintain 75% to 100% of original length after exiting the chamber.

16. The method of claim 13, wherein the nucleic acids maintain 90% to 100% of original length after exiting the chamber.

17. The method of claim 13, wherein the nucleic acids include DNA having lengths of about 1 megabase in length after exiting the chamber.

18. The method of claim 1, further comprising:

conveying flow with agents positioned in central streamlines from the chamber prior to removing flow from about the central streamlines.

19. The method of claim 18, wherein conveying comprises conveying the fluid of a distance greater than about 10 cm.

20. The method of claim 18, wherein conveying comprises conveying the fluid of a distance greater than about 20 cm.

21. The method of claim 20, wherein agents are conveyed without dispersing more than 2 cm in a direction of flow.

22. The method of claim 21, wherein agents are conveyed without dispersing more than 2 cm in a direction of flow.

23. The composition of claim 22, wherein the composition comprises at least 100 ng of nucleic acids.

24. The composition of claim 22, wherein the nucleic acids are in a concentration of about 3 to about 6 ng per uL.

22-1. A composition comprising isolated nucleic acids varying in length from 0.1 megabases to about 1 megabase in a fluid having a volume between about 30 uL to about 40 uL.

25. A method positioning agents in a fluid flow, the method comprising:

providing a substrate;

flowing a first fluid containing agents through the substrate;

flowing a second fluid flow through the substrate, the second fluid flow at least partially surrounding the first fluid flow to direct the first fluid flow and any agents contained therein toward a central portion of the substrate;

positioning the agents on the central portion of the substrate; and

reversing flow through substrate to move any agents positioned on the substrate away from the substrate in central streamlines of fluid moving away from the substrate.