DNA isolation method

• US 20050019819A1
• Filed: 08/23/2004
• Published: 01/27/2005
• Est. Priority Date: 10/28/1999
• Status: Active Grant

• Current Assignee: Gyros Patent AB (Mesa Laboratories Inc.)
• Original Assignee: Gyros Patent AB (Mesa Laboratories Inc.)
• Inventors: Kenrick, Michael Kenneth, Tooke, Nigel Eric, Thomas, Philip Landeg
• Granted Patent: US 6,992,181 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• 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

Disclosed is a method and apparatus for the isolation of DNA or cell nuclei or a mixture thereof from cell samples in a CD device. The method includes treating a suspension of whole cells with a lysis reagent so as to lyse the cytoplasmic membrane and at least some of the nuclear membranes, and introducing the lysate into micro-channels of a microfabricated apparatus in which each of the micro-channels is provided with a barrier disposed in the channel to impede the passage or flow of DNA and cell nuclei while allowing the passage of liquid through the micro-channel so that a mesh comprising DNA is formed in the channel.

Claims

1-22. -22. (canceled).

23. A method of isolating DNA and/or cell nuclei in a microfabricated apparatus that contains a plurality of micro-channels, the method comprising the steps of:

(i) providing a cell lysate; and

(ii) forming in one or more of the micro-channels a mesh of nucleic acid of the lysate, wherein said mesh acts as a barrier to DNA and/or cell nuclei.

24. The method of claim 23, wherein multiple cell lysates are treated in parallel within the microfabricated device.

25. A method of isolating DNA and/or cell nuclei in a microfabricated apparatus that comprises at least one micro-channel having an inlet channel and an outlet channel, said method comprising the step of forming a mesh from a cell sample in at least one micro-channel, wherein said mesh impedes the passage or flow of DNA and/or cell nuclei while allowing passage of liquid through said micro-channel.

26. The method of claim 25, wherein the microfabricated apparatus is further defined as a rotatable disc.

27. The method of claim 25, wherein said micro-channel further comprises a reaction chamber for performing assays or processing DNA.

28. The method of claim 27, wherein said assays or processing are polymerase chain reaction or restriction enzyme digestion.

29. The method of claim 25, wherein said micro-channel comprises a means to impede the passage or flow of DNA and/or cell nuclei while allowing passage of liquid through said micro-channel.

30. The method of claim 29, wherein said means to impede the passage or flow of DNA and/or cell nuclei while allowing passage of liquid through said micro-channel is a barrier.

31. The method of claim 30, wherein said barrier comprises a bead.

32. The method of claim 31, wherein said bead is selected from the group consisting of a polystyrene bead, a styrene acrylate co-polymer bead, a polystyrene cross-linked with divinylbenzene bead, a polyvinyltoluene bead, a polymethacrylate bead, a polycarbonate bead, a polysaccharide bead, a metal oxide bead, a glass bead and a carbon bead.

33. The method of claim 25, wherein said barrier comprises raised structures disposed in the micro-channel.

34. The method of claim 33, wherein said raised structures are disposed on the base portion of said micro-channel and are moulded to form pillars.

35. The method of claim 25, wherein said cell sample is plant cells or animal cells.

36. The method of claim 25, wherein said nuclei captured by the mesh are further treated to release DNA.