Process for microfabrication of an integrated PCR-CE device and products produced by the same
First Claim
1. An integrated microvolume device for performing polymerase chain reaction and electrophoresis, the device comprising:
- at least one reaction chamber, the reaction chamber comprisingtwo planar members coupled together at at least one point, wherein one planar member has a thickness of up to about 1 mm and is thicker than the other planar member;
a cavity disposed within the planar members;
at least one aperture in communication with the cavity to permit the introduction and withdrawal of fluids therefrom;
a thin film metal heater disposed within the cavity; and
a thermocouple having a temperature-sensing junction disposed adjacent to the cavity;
an electrophoresis capillary; and
channels in fluid communication with the cavity and the electrophoresis capillary.
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Abstract
A fully integrated monolithic small volume PCR-CE device in glass, or the like materials, is fabricated using thin film metal heaters and thermocouples to thermally cycle sub-microliter PCR volumes. Successful amplification of a PCR fragment is demonstrated on a PCR-CE chip. The process utilizes a linear polyacrylamide surface coating coupled with addition of BSA to the amplification buffer was necessary to obtain amplification efficiencies comparable to a positive control. The micro-reactor reduced significantly the time required for amplification and the reaction volume was in the sub-microlitre regime. Likewise addressed are the known problems connected with reliable microfabrication of metal coatings and the insulating layers required to shield these layers from the PCR reaction mix, and the longstanding unresolved issue of exposed metal regions in the PCR-CE chip resulting in electrolysis of water and bubble formation whenever a voltage is applied. The instant teachings employ external heaters and thermocouples and, as such, have alleviated many of these problems. Heaters and thermocouples may still be thin film deposited after chip bonding allowing for easy scale-up to multichannel devices. In addition, direct deposition of these chip components insures good thermal contact with the PCR reactor.
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Citations
29 Claims
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1. An integrated microvolume device for performing polymerase chain reaction and electrophoresis, the device comprising:
- at least one reaction chamber, the reaction chamber comprising
two planar members coupled together at at least one point, wherein one planar member has a thickness of up to about 1 mm and is thicker than the other planar member;
a cavity disposed within the planar members;
at least one aperture in communication with the cavity to permit the introduction and withdrawal of fluids therefrom;
a thin film metal heater disposed within the cavity; and
a thermocouple having a temperature-sensing junction disposed adjacent to the cavity;
an electrophoresis capillary; and
channels in fluid communication with the cavity and the electrophoresis capillary. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
- at least one reaction chamber, the reaction chamber comprising
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18. An integrated microvolume device for performing polymerase chain reaction and electrophoresis, the device comprising at least one reaction chamber, the reaction chamber comprising
two generally planar members coupled together in an overlapping relationship, each planar member having a cavity defining a chamber therebetween, wherein one planar member has a thickness of up to about 1 mm and is thicker than the other planar member, and wherein wherein at least one of the planar members has an elongated aperture in flow communication with the chamber to permit the introduction and withdrawal of fluids therefrom; -
a thin film heater disposed within the chamber; and
a thermocouple having a temperature-sensing junction disposed in one of the planar members, being positioned so as to sense the temperature of a reaction occurring within the chamber an electrophoresis capillary; and
channels in fluid communication with the chamber and the electrophoresis capillary. - View Dependent Claims (19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29)
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Specification