Crude biological derivatives competent for nucleic acid detection
OwnerBrittan L. Pasloskeinventor
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Abstract
The invention relates to methods for the detection of a specific sequence of RNA in a cell or tissue sample. The invention also relates to methods to enzymatically manipulate the RNA in a crude cell lysate in a number of applications.
Claims70
A method comprising:
- obtaining at least one biological unit containing RNA;
- obtaining at least one catabolic enzyme;
- preparing an admixture of the biological unit and the catabolic enzyme;
- and
- incubating the admixture under conditions where the catabolic enzyme is active.
The method of claim 28, wherein the RNase inhibitor is a non-proteinaceous RNase inhibitor
A method for producing cDNA from one or more biological units comprising:
- obtaining at least one biological unit;
- obtaining at least one catabolic enzyme;
- preparing an admixture of the biological unit and the catabolic enzyme;
- and
- incubating the admixture at a temperature where the catabolic enzyme is active and with reverse transcriptase under conditions to allow reverse transcription.
A kit for producing cDNA from a biological unit, comprising, in a suitable container:
- a buffer;
- and
- a catabolic enzyme.
A kit for producing cDNA from a biological unit comprising, in one or more suitable container(s):
- a biological unit lysis buffer;
- a deoxyribonuclease;
- an RNase inhibitor;
- a reverse transcription buffer;
- reverse transcriptase;
- dNTPs;
- and
- an Armored RNA® control.
A Cell Lysis Buffer comprising a catabolic enzyme, 1 mM CaCl2, 3 mM MgCl2, 1 mM EDTA, 1% Triton X100, and 10 mM Tris pH 7.5.
Dependencies are inferred from the claim text.