DNA isolation method

• US 6,992,181 B2
• Filed: 08/23/2004
• Issued: 01/31/2006
• Est. Priority Date: 10/28/1999
• Status: Expired due to Fees

• Current Assignee: Gyros Patent AB (Mesa Laboratories Inc.)
• Original Assignee: Gyros AB (Mesa Laboratories Inc.)
• Inventors: Tooke, Nigel Eric, Kenrick, Michael Kenneth, Thomas, Philip Landeg
• Primary Examiner(s): KETTER, JAMES S
• Application Number:
  US10/924,151
• Publication Number:
  US 20050019819A1
• Time in Patent Office: 526 Days
• Field of Search: 536/25.4, 435/306.1, 435/317.1
• US Class Current: 536/25.4
• 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. 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, wherein multiple cell lysates are treated in parallel within the microfabricated device.

2. 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, and wherein multiple samples are treated in parallel within the microfabricated device.

3. The method of claim 2, wherein the microfabricated apparatus is further defined as a rotatable disc.

4. The method of claim 2, wherein said micro-channel further comprises a reaction chamber for performing assays or processing DNA.

5. The method of claim 4, wherein said assays or processing are polymerase chain reaction or restriction enzyme digestion.

6. The method of claim 2, 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.

7. The method of claim 6, 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.

8. The method of claim 7, wherein said barrier comprises a bead.

9. The method of claim 8, 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.

10. The method of claim 2, wherein said barrier comprises raised structures disposed in the micro-channel.

11. The method of claim 10, wherein said raised structures are disposed on the base portion of said micro-channel and are moulded to form pillars.

12. The method of claim 2, wherein said cell sample is plant cells or animal cells.

13. The method of claim 2, wherein said nuclei captured by the mesh are further treated to release DNA.