Multiplexed polymerase chain reaction in micropipette format
View Patent ↗A system may include a horizontal actuator to move a tray, to which a microwell plate and a microfluidic chip may be coupled. The system may include a vertical actuator to move a support arm, to which a plurality of pipettes or pipette tips may be coupled. The system may include a rotational actuator to move an angle bracket, to which a magnet may be coupled. The system may include a heater, through which the pipettes may extend. The system may include a pump to control the flow of fluids through the pipettes. Disclosed methods include performing PCR within the described system.
1. A method, comprising:
operating a pump to draw a biological sample from a well of a well plate into a pipette through a pipette tip of the pipette;
operating a first actuator to move a first valve to a closed position to seal the biological sample within the pipette from the pipette tip;
operating the pump to apply a positive gage pressure to the biological sample within the pipette;
operating a second actuator to move a second valve to a closed position to seal the biological sample within the pipette from the pump; and
operating a heater to heat the biological sample within the pipette, wherein a polymerase chain reaction occurs within the heated biological sample within the pipette between the first valve and the second valve.
2. The method of claim 1 wherein the positive gage pressure is at least 7 psi above atmospheric pressure.
3. The method of claim 1 , further comprising:
operating the second actuator to move the second valve to an open position;
operating the pump to release the positive gage pressure;
operating the first actuator to move the first valve to an open position; and
operating the pump to dispense the biological sample through the pipette tip of the pipette.
4. The method of claim 3 wherein operating the second actuator to move the second valve to an open position takes at least about five seconds.
5. The method of claim 3 wherein operating the first actuator to move the first valve to an open position takes at least about five seconds.
6. A method, comprising:
operating a pump to draw a biological sample from a well of a well plate into a first pipette conduit through a first pipette tip;
operating a first actuator to move a first valve to a closed position to seal the biological sample within the first pipette conduit from the first pipette tip;
operating the pump to apply a positive gage pressure to the biological sample within the first pipette conduit;
operating a second actuator to move a second valve to a closed position to seal the biological sample within the first pipette conduit from the pump; and
operating a heater to heat the biological sample within the first pipette conduit, wherein a chemical reaction occurs within the heated biological sample within the first pipette conduit between the first valve and the second valve.
7. The method of claim 6 , further comprising:
operating the second actuator to move the second valve to an open position;
operating the pump to release the positive gage pressure;
operating the first actuator to move the first valve to an open position; and
operating the pump to draw the biological sample through a three-way connector.
8. The method of claim 7 wherein operating the second actuator to move the second valve to an open position takes at least about five seconds.
9. The method of claim 7 wherein operating the first actuator to move the first valve to an open position takes at least about five seconds.
10. The method of claim 7 , further comprising:
operating the first actuator to move the first valve to the closed position or operating the second actuator to move the second valve to the closed position; and
operating the pump to push the biological sample through the three-way connector into a second pipette conduit.
11. The method of claim 10 , further comprising:
operating the pump to push the biological sample through the second pipette conduit; and
operating the pump to dispense the biological sample through a second pipette tip coupled to the second pipette conduit.
12. The method of claim 11 wherein the chemical reaction is a polymerase chain reaction.
13. The method of claim 10 , further comprising:
operating a third actuator to move a third valve to a closed position to seal the biological sample within the second pipette conduit from a second pipette tip coupled to the second pipette conduit;
operating the pump to apply a second positive gage pressure to the biological sample within the second pipette conduit;
operating a fourth actuator to move a fourth valve to a closed position to seal the biological sample within the second pipette conduit from the pump; and
operating a heater to heat the biological sample within the second pipette conduit, wherein a second chemical reaction occurs within the heated biological sample within the second pipette conduit between the third valve and the fourth valve.
14. The method of claim 13 , further comprising:
operating the fourth actuator to move the fourth valve to an open position;
operating the pump to release the second positive gage pressure;
operating the third actuator to move the third valve to an open position; and
operating the pump to dispense the biological sample through the second pipette tip.
15. The method of claim 14 wherein operating the fourth actuator to move the fourth valve to an open position takes at least about five seconds.
16. The method of claim 14 wherein operating the third actuator to move the third valve to an open position takes at least about five seconds.
17. The method of claim 13 wherein the second chemical reaction is a polymerase chain reaction.
18. The method of claim 13 wherein the second positive gage pressure is at least 7 psi above atmospheric pressure.
19. The method of claim 13 wherein operating the fourth actuator to move the fourth valve to a closed position seals the biological sample within the second pipette conduit from the three-way connector.