Method and formulation for lyophilizing cultured human cells to preserve RNA and DNA contained in cells for use in molecular biology experiments
AssigneeBio-Rad Laboratories, Inc.
InventorsChing-I Patsy Lin, Robert Bruce Wallace, Jeffrey Cossman, Cynthia French
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Abstract
The present invention relates to methods for lyophilizing eukaryotic cells and isolating intact nucleic acids from such cells, and related kits for using the same.
Claims30
A method for long term preservation of nucleic acid combined within eukaryotic cells the method comprising:
- i. lyophilizing an aqueous solution of intact eukaryotic cells where said solution is isotonic to the cells and is nuclease free;
- and,
- ii. maintaining the lyophilized cells under sealed conditions sufficient to avoid contact with atmospheric humidity.
A method for isolating intact nucleic acid from lyophilized cells comprising:
- i. lyophilizing an aqueous solution of living eukaryotic cells where said solution is isotonic to the cells and is nuclease free;
- ii. maintaining the lyophilized cells under sealed conditions for at least 30 days;
- iii. unsealing the cells;
- iv. denaturing the cellular proteins to create a mixture of intact nucleic acid and denatured cellular proteins;
- and,
- v. isolating intact nucleic acid from the mixture of denatured cellular proteins and nucleic acid with the proviso that the cells are not revived.
A collection of standardized, sealed vials containing lyophilized eukaryotic cells for use as controls in diagnostic assays wherein the cells after 4 weeks at −20° C. have more than 50% of their 18S rRNA intact as measured by gel electrophoresis.
A nucleic acid hybridization assay kit comprising lyophilized eukaryotic cells.
Dependencies are inferred from the claim text.