Nucleic acid processing kits and devices

摘要

Biological samples containing nucleic acids, RNA and DNA, are freed from bound proteins by incubation with a chaotropic agent such as a guanidium salt, and the mixture is readied for further processing by dilution of such agent to a concentration below 0.05 M without physical isolation of RNA and DNA from one another or from other components of the reaction mixture. Methods include such preparation and further processing, such as amplification and detection, which may be performed in a single container. Chaotropic agent may be supplied as a dried reagent adhered to a container. Kits may include such reagents, alone or with amplification reagents.

主权项

1. A method for enzymatically synthesizing cDNA from a biological sample, comprising:
(a) combining said biological sample with a disruption reagent in a container to produce a mixture comprising at least 2M chaotropic agent; (b) diluting said mixture with at least one aqueous reagent to reduce the concentration of said chaotropic agent to less than 0.05 M; and (c) incubating at least a portion of said mixture with reverse transcriptase.