Microfabricated injector and capillary array assembly for high-resolution and high throughput separation

• US 6,533,914 B1
• Filed: 06/27/2000
• Issued: 03/18/2003
• Est. Priority Date: 07/08/1999
• Status: Expired due to Fees

• Current Assignee: Key Safety Systems, Inc. (Ningbo Junsheng Investment Group Co., Ltd.)
• Original Assignee: Shaorong Liu
• Inventors: Liu, Shaorong
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): BROWN, JENNINE M
• Application Number:
  US09/604,861
• Time in Patent Office: 994 Days
• Field of Search: 204/600, 204/601, 204/451, 204/452, 204/603
• US Class Current: 204/601
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0838 Capillaries
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/563 Joints or fittings in general F16L; Separable fluid transfer means to transfer fluids between at least two containers, e.g. connectors
• CPC Class Codes: G01N 27/44743 Introducing samples
• CPC Class Codes: G01N 27/44782 of a plurality of samples
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027

Abstract

The present invention concerns methods and apparatus for the high resolution, high output electrophoretic separation of molecules. In preferred embodiments, the methods and apparatus are of use for DNA sequencing. The apparatus comprises a hybrid device, comprising a microfabricated chip injector attached to an array of one or more capillaries. The chip injector is designed with incorporation and injector channels that precisely match the capillaries, to minimize or eliminate dead volume in the system. DNA sequencing runs of over 700 bases, with a run time of less than one hour, may be accomplished with the methods and apparatus disclosed herein.

Claims

1. A hybrid apparatus for high throughput, high resolution electrophoretic separation comprising a microfabricated chip injector attached to one or more capillaries, each capillary connected in a straight line with an injection channel in the chip injector.

2. The apparatus of claim 1, wherein the microfabricated chip injector comprises:

a) one or more connection channels that match the outside shape of the capillary; and

b) one or more injector channels that match the inside shape of the capillary.

3. The apparatus of claim 2, wherein the capillary is inserted into the connection channel until the injection channel is flush with a channel inside the capillary, resulting in an assembly having minimal dead volume.

4. The apparatus of claim 3, wherein each injection channel is connected by a first cross channel to a sample reservoir, by a second cross channel to a waste reservoir, and by a third cross channel to a cathode reservoir.

5. The apparatus of claim 3, wherein the injection channel is smoothly connected to the e channel inside the capillary.

6. The apparatus of claim 3, wherein the apparatus comprises at least 10 capillaries.

7. The apparatus of claim 6, wherein the apparatus comprises at least 100 capillaries.

8. The apparatus of claim 7, wherein the apparatus comprises at least 200 capillaries.

9. The apparatus of claim 8, wherein the apparatus comprises at least 300 capillaries.

10. The apparatus of claim 9, wherein the apparatus comprises at least 400 capillaries.

11. The apparatus of claim 10, wherein the apparatus comprises at least 500 capillaries.

12. The apparatus of claim 11, wherein the apparatus comprises at least 700 capillaries.

13. The apparatus of claim 12, wherein the apparatus comprises at least 900 capillaries.

14. The apparatus of claim 3, wherein the apparatus is capable of performing DNA sequence analysis of at least 500 bases of DNA sequence.

15. The apparatus of claim 14, wherein the apparatus is capable of performing DNA sequence analysis of at least 800 bases of DNA sequence.

16. The apparatus of claim 15, wherein the apparatus is capable of performing DNA sequence analysis of between 800 and 1000 bases of DNA sequence.

17. The apparatus of claim 15, wherein the apparatus is capable of performing DNA sequence analysis of greater than 1000 bases of DNA sequence.

18. The apparatus of claim 3, wherein the apparatus is designed to operate with a sample volume of 5.0 μ

l or less.

19. The apparatus of claim 3, further comprising a rotary scanner.

20. A method of sequencing DNA, comprising:

a) performing a Sanger dideoxy reaction with a sample of DNA to be sequenced;

b) loading the products of said reaction into one or more sample reservoirs of the apparatus of claim 3;

c) performing an electrophoretic separation of said products;

d) detecting said separated products; and

e) analyzing said detected products to provide DNA sequence data.

21. The method of claim 20, wherein said reaction products are fluorescently labeled.

22. The method of claim 21, further comprising four color DNA sequencing.