DNA analyzer

• US 9,656,261 B2
• Filed: 03/04/2011
• Issued: 05/23/2017
• Est. Priority Date: 06/04/2009
• Status: Active Grant

• Current Assignee: Leidos Innovations Technology, Inc., Microgem NZ Limited, University of Virginia Patent Foundation (University of Virginia)
• Original Assignee: Leidos Innovations Technology, Inc., University of Virginia Patent Foundation (University of Virginia), Zygem Corporation Ltd.
• Inventors: Bienvenue, Joan M, Pettit, John W, Norris, Jessica V, Marchiarullo, Daniel J, Bell, H. Randall, Landers, James P, Scott, Orion N, Leslie, Daniel C
• Primary Examiner(s): MUMMERT, STEPHANIE KANE
• Application Number:
  US13/064,094
• Publication Number:
  US 20110229898A1
• Time in Patent Office: 2,272 Days
• Field of Search: None
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: G01N 2021/4186 Phase modulation imaging
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 29/32 Arrangements for suppressing undesired influences, e.g. temperature or pressure variations , compensating for signal noise

Abstract

Aspects of the disclosure provide a microfluidic chip to facilitate DNA analysis. The microfluidic chip includes a first domain configured for polymerase chain reaction (PCR) amplification of DNA fragments, a dilution domain coupled to the first domain to dilute a PCR mixture received from the first domain, and a second domain that is coupled to the dilution domain so as to receive the amplified DNA fragments. The second domain includes a separation channel that is configured to perform electrophoretic separation of the amplified DNA fragments. In addition, the disclosure provides a DNA analyzer to act on the microfluidic chip to perform an integrated single chip DNA analysis.

Claims

1. A microfluidic chip, comprising:

an inlet configured to receive a template DNA extracted from a sample acceptor that is external of the microfluidic chip and is fluidically coupled to the microfluidic chip on a cartridge;

a first domain configured for polymerase chain reaction (PCR) amplification of DNA fragments, the first domain including a thermal-coupler reservoir coupled with a PCR reservoir for PCR amplification, thermal cycles in the PCR reservoir being induced based on temperature measured from the thermal-coupler reservoir;

a dilution domain having a reservoir that is fluidically coupled to the first domain and is configured to dilute a PCR mixture received from the first domain with a dilutant in the reservoir; and

a second domain that is fluidically coupled to the dilution domain so as to receive the amplified DNA fragments, the second domain including a separation channel that is configured to perform electrophoretic separation of the amplified DNA fragments.

2. The microfluidic chip of claim 1, wherein:

the PCR reservoir is configured to receive a template DNA and reagents, and amplify the DNA fragments based on the template DNA and the reagents.

3. The microfluidic chip of claim 2, further comprising:

a plurality of inlets configured to input the template DNA and the reagents into the microfluidic chip.

4. The microfluidic chip of claim 1, wherein the second domain comprises:

an injection channel configured to inject the amplified DNA fragments into the separation channel by electro-kinetic injection.

5. The microfluidic chip of claim 1, further comprising:

a plurality of electrode reservoirs for applying an electric field over the separation channel.

6. The microfluidic chip of claim 1, wherein in the dilution domain, the PCR mixture and the dilutant are mixed according to a ratio from 1:

5 to 1;

20.

7. A cartridge, comprising:

a sample acceptor external to and fluidically coupled to a microfluidic chip on the cartridge,

the sample acceptor configured to extract a template DNA and the microfluidic chip having a first domain and a second domain coupled to the first domain, wherein the first domain is configured to perform polymerase chain reaction (PCR) amplification of DNA fragments based on the template DNA, and the second domain has a separation channel that is configured to perform electrophoretic separation of the amplified DNA fragments, the first domain includes a thermal-coupler reservoir coupled with a PCR reservoir for PCR amplification, thermal cycles in the PCR reservoir are induced based on temperature measured from the thermal-coupler reservoir.

8. The cartridge of claim 7, further comprising:

a reagent carrier configured to carry reagents for the PCR amplification, and solutions for the electrophoretic separation.

9. The cartridge of claim 7, wherein the sample acceptor is configured to extract the template DNA by at least one of a silica solid phase extraction, and a liquid phase enzymatic DNA isolation.

10. The cartridge of claim 7, wherein the sample acceptor comprises:

a well having a liquid phase mixture to extract the template DNA.

11. The cartridge of claim 10, wherein the liquid phase mixture is sealed in the well before the extraction.

12. The cartridge of claim 8, wherein:

the PCR reservoir is configured to receive the extracted template DNA and the reagents, and amplify the DNA fragments based on the template DNA and the reagents.

13. The cartridge of claim 8, wherein the microfluidic chip further comprises:

a plurality of inlets configured to input the template DNA and the reagents kits into the microfluidic chip.

14. The cartridge of claim 7, wherein the second domain of the microfluidic chip comprises:

an injection channel configured to inject the amplified DNA fragments into the separation channel by electro-kinetic injection.

15. The cartridge of claim 7, wherein the microfluidic chip further comprises:

a plurality of electrode reservoirs for applying an electric field over the separation channel.

16. The cartridge of claim 7, wherein the microfluidic chip further comprises:

a waste outlet to drain liquid out of the microfluidic chip.

17. The cartridge of claim 7, wherein the microfluidic chip further comprises:

a dilution domain coupled to the first domain and the second domain, the dilution domain diluting a PCR mixture received from the first domain with a dilutant, and providing the diluted PCR mixture to the second domain.

18. The cartridge of claim 17, wherein in the dilution domain, the PCR mixture and the dilutant are mixed according to a ratio from 1:

5 to 1;

20.

19. The cartridge of claim 10, wherein the liquid phase mixture is capable of performing enzymatic digestion of proteins and other cellular interferences, with the exception of DNA.