In situ PCR amplification system
First Claim
1. A containment system for in situ PCR amplification of a target nucleic acid sequence comprising:
- a glass microscope slide;
a sample containing said target sequence mounted on said slide;
a reagent mixture on said sample;
a resilient and compliant cover member on said slide over said sample and said reagent mixture; and
a retaining assembly mechanically fastened to said slide and fastened to said cover member, said assembly including a seal member around a rim portion of said cover member, said assembly being operable to retain said sample and said mixture between said cover member and said slide and seal said cover member against said slide.
9 Assignments
0 Petitions
Accused Products
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.
112 Citations
54 Claims
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1. A containment system for in situ PCR amplification of a target nucleic acid sequence comprising:
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a glass microscope slide; a sample containing said target sequence mounted on said slide; a reagent mixture on said sample; a resilient and compliant cover member on said slide over said sample and said reagent mixture; and a retaining assembly mechanically fastened to said slide and fastened to said cover member, said assembly including a seal member around a rim portion of said cover member, said assembly being operable to retain said sample and said mixture between said cover member and said slide and seal said cover member against said slide. - View Dependent Claims (15)
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- 2. The system according to claim I wherein said retaining assembly comprises at least one clip means for removably retaining said cover member against said slide.
- 16. A retaining assembly for retaining a fluid against a glass slide comprising a resilient and compliant cover member, an annular seal member around at least a portion of said cover member and at least one clip means for removably retaining said cover and seal members against said slide.
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31. A system for in situ PCR amplification of a nucleic acid sequence contained in a sample comprising:
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a glass microscope slide having said sample mounted thereon; a reagent mixture on said sample; a resilient and compliant cover member over said sample and said reagent mixture on said slide; a retaining assembly fastened to said slide and to said cover member operable to retain and seal said sample and said mixture between said cover member and said slide; and a thermal cycler having a temperature controlled block adapted to receive and hold said microscope slide against a portion of said block so that the temperature of said sample on said slide follows the temperature of said temperature controlled block. - View Dependent Claims (32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47)
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- 48. A retaining assembly for retaining a fluid against a glass slide comprising a resilient and compliant cover member, an annular seal member around at least a portion of said cover member and at least one clip means for removably retaining said cover and seal members against said slide, said cover member having an annular ridge between said portion of said cover member and a central portion of said cover member.
Specification