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Abstract

A complete in situ PCR system for amplification of nucleic acids contained in a prepared cell or tissue sample. The containment system for the PCR sample comprises a glass microscope slide, a specimen sample containing the target nucleic acid sequence mounted on the slide, a flexible plastic cover over the sample, and a retaining assembly fastened to the slide and to the cover to retain and seal a reaction mixture against the sample during thermal cycling. The retaining assembly includes a rigid ring on a rim portion of the cover, a cross beam having spaced parallel rails joined by opposite flat ends, and a pair of clips which are pressed over the ends and opposite sides of the slide to fasten the cross beam and cover to the slide. In one form the retaining assembly includes a flexible compliant cover member, which is permeable to gas but non-permeable to liquid, for forming a closed liquid space adjacent a flat surface, a cross beam member enclosing the cover member to form a closed gas space therebetween, the periphery of the liquid space and of the gas space being sealed, and a pressure differential being established whereby the pressure in the liquid space is greater than the pressure in the gas space.

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First Claim