Crude biological derivatives competent for nucleic acid detection
AssigneeLife Technologies Corporation
InventorsBrittan L. Pasloske, Xingwang Fang, Quoc Hoang
Life of this patent
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Abstract
The invention relates generally to the fields of making biological unit lysates or admixtures of body fluids and of RNA analysis. More specifically, it relates to direct methods for the detection of a specific sequence of RNA in a biological unit, for example a virus, cell or tissue sample, or a body fluid, for example saliva, sputum, blood plasma, etc. More generally, the invention may be used to enzymatically manipulate and protect the RNA in lysate or bodily fluids for a number of applications.
Claims10
A method comprising:
- obtaining at least one biological unit containing RNA or sample comprising RNA not comprised in a biological unit;
- obtaining a low pH buffer;
- mixing the biological unit and the buffer to prepare a low pH lysate or mixing the sample and the buffer to prepare a low pH admixture wherein the low pH lysate or admixture has a pH of less than 5;
- and
- mixing at least a portion of the lysate or admixture with a composition comprising enzyme using RNA as a substrate to form a reaction mixture.
A kit for assaying RNA in a biological unit or sample comprising, in one or more suitable container(s):
- a low pH buffer, a high pH buffer, or a buffer that precipitates RNA;
- a reverse transcription buffer;
- reverse transcriptase;
- and
- dNTPs.
A method comprising:
- obtaining at least one biological unit containing RNA or sample comprising RNA not comprised in a biological unit;
- obtaining a high pH buffer;
- mixing the biological unit and the buffer to prepare a high pH lysate or mixing the sample and the buffer to prepare a high pH admixture;
- and
- mixing at least a portion of the lysate or admixture with a composition comprising an enzyme using RNA as a substrate to form a reaction mixture.
Dependencies are inferred from the claim text.