Use of centrifugation to prepare a retractable seal over reagent

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or...

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435 6, 435 71, 435 41, 435 89, 435 911, 435 914, 435 9141, 435174, 435177, 435182, 436528, 436535, 530812, 530817, C12Q 100, C12Q 168, G01N 3353, C12P 1934

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059687290

ABSTRACT:
A method is provided using centrifugation to prepare a seal of solidified wax, grease or polymer mix over an aqueous reagent in a reaction container such that the reagent is separated from contact with the atmosphere. The amount of solidified wax, grease or polymer mix is not sufficient when melted to a liquid to separate the reagent from contact with the atmosphere under gravity. A reagent and solidified wax, grease or polymer mix are combined in a container. During centrifugation and heating, the solidified wax, grease or polymer mix melts to a liquid, and centrifuging causes the liquid to form over the reagent a layer that completely separates the reagent from the atmosphere. As centrifugation continues, the liquid is cooled and solidified to form the seal. Additional reagents are preferably added on top of the seal such that when the container is heated and the seal melted the upper and lower reagents mix for reaction. Preferred use of sealing reagents by this method is in polymerase chain reaction (PCR), reverse transcriptase reactions, nucleic acid sequencing, and colorimetric, fluorometric or chemiluminescent labeled immunoassays.

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