Integrated microvolume device
First Claim
1. In a fully integrated glassine monolithic microvolume PCR-CE device, the improvement which comprises:
- utitilizing thin film metal heating means to maintain a predetermined range of temperature for desired denaturations; and
employing thermocoupling means for assuring requisite cycling of sub-microliter PCR volumes.
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Accused Products
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-microliter 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.
229 Citations
20 Claims
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1. In a fully integrated glassine monolithic microvolume PCR-CE device,
the improvement which comprises: -
utitilizing thin film metal heating means to maintain a predetermined range of temperature for desired denaturations; and
employing thermocoupling means for assuring requisite cycling of sub-microliter PCR volumes. - View Dependent Claims (2, 3, 4, 14, 15, 16, 20)
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5. An integrated monolithic PCR reactor coupled to a microcapillary electrophoresis system including an electrophoresis column, the apparatus comprising in combination:
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a reaction cavity microfabricated within two-bonded substrates defining a space therebetween;
a plurality of apertures functioning as access means for introducing fluids into said reaction cavity;
exit channel means for connecting the reactor to the electrophoresis system and for injecting material onto the electrophoresis column incorporated therein;
a heating means for regulating temperature disposed on the outside of one of said bonded substrates; and
,a thermocouple disposed in a least one position selected from the group consisting of within the space defined by said reaction cavity and between said heater and said bonded substrate. - View Dependent Claims (6, 7, 8, 9, 10, 11, 12, 13, 17, 18, 19)
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Specification