Process for microfabrication of an integrated PCR-CE device and products produced by the same

DETAILED DESCRIPTION A submicroliter PCR type of a reaction chamber is taught for the amplification of specific diagnostic targets using PCR, among other things.
Subject amplicons are then directly injected into microfabricated CE channels for fragment size analysis, or use with related biomolecular assays.
According to a preferred embodiment, PCR chambers and CE channels are etched into a thickened glass substrate using chemical etching.
A thin (for example 0.
20 mm) glass substrate is bonded to the etched surface defining the chambers and channels.
The thin substrate utilizes a thermocouple or platinum resistance temperature sensing device on its interior or exterior surface in combination with a platinum or Peltier heater on the external surface for driving thermal cycling.
Briefly stated, 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 to obtain amplification efficiencies comparable to a positive control.
The micro-reactor reduces significantly the time required for amplification and the reaction volume within the context of a 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 itself, 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, for example, a PCR reactor