Methods for the recovery of nucleic acids from reaction mixtures

Details Methods for the recovery of nucleic acids from reaction mixtures Methods for the recovery of nucleic acids from reaction mixtures

1991-07-03

1995-06-06

Robinson, Douglas W.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435184, 435195, 935 77, 935 19, 536 254, 536 2541, 536 2542, C12N 922, C12N 999

Patent

active

054222419

ABSTRACT:
A method for the recovery of nucleic acids from a reaction mixture, such as nuclease protection assays, comprising the use of one reagent containing a) nuclease for digesting single-stranded RNA and b) a nucleic acid precipitating carrier agent, (sheared DNA or linear acrylamide). A second reagent contains a chaotropic agent (a guanidinium salt) for inactivating said nucleases and an alcohol (ethanol or isopropanol) for the simultaneous inactivation of the nucleases and the precipitation of the nucleic acids without the need for protease digestion or organic extraction.

REFERENCES:
patent: 4843155 (1989-06-01), Chomczynski
Cox, R. A.: Methods in Enzymology 12(B):121-129, 1968.
Gilman, M., "Ribonuclease Protection Assay," Current Protocols in Molecular Biology, Wiley Interscience, vol. 1, Suppl. 7, pp. 4.7.1-4.7.8, 1989.
Sambrook et al., "Mapping of RNA with Ribonuclease and Radiolabeled RNA Probes," Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbour Laboratroy Press, pp. 7.71-7.77, 1989.
Chomczynski, P. and Sacchi, N., "Single-Step Method of RNA Isolation by Acid Guanidinium Thiocyanate-Phenol-Chloroform Extraction," Anal. Biochem., 162:156-159, 1987.
"Ambion Catalog #1400: RPA" Ribonuclease Protection Assay Kit Instruction Manual.

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