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US 4,900,677 AGranted
Process for rapid isolation of high molecular weight DNA
AssigneeE. I. Du Pont de Nemours and Company
InventorPeter L. Hewitt
Life of this patent
Estimating the term…
Abstract
A procedure for isolating high molecular weight nucleic acids utilizing a mixture of lytic enzymes and a chaotropic agent to complete protein denaturation and dissociation from nucleic acids is provided. The nucleic acids so obtained are useful for restriction enzyme analysis and DNA probe hybridization.
Claims8
A method of isolating nucleic acids from their source organism comprising the steps of:
- (A) forming a suspension of said organism containing or suspected of containing the desired nucleic acid;
- (B) treating said organism with a cocktail of lytic enzymes;
- (C) treating said organism with surfactant prior to, simultaneously with or subsequent to step (B);
- (D) degrading unwanted classes of nucleic acids by treatment with nucleases specific for the unwanted nucleic acids;
- then
- (E) degrading proteins by digestion with at least one broadly active protease;
- (F) denaturing remaining proteins and dissociating them from the nucleic acid by adding at least one chaotropic agent;
- and then
- (G) dialyzing and concentrating the nucleic acid.
6 more elements
A method of isolating DNA from cells or viruses comprising the steps of:
- (A) forming a suspension of said cells or viruses containing or suspected of containing the desired DNA;
- (B) treating said cells or viruses with a mixture of lytic enzymes comprising lysozyme, endo-N-acetylmuraminidase and achromopeptidase;
- (C) treating said cells or viruses with sodium dodecyl sulfate;
- (D) degrading RNA from said cells or viruses with RNases;
- then
- (E) degrading proteins by digestion with Proteinase K;
- (F) denaturing remaining proteins and dissociating them from DNA by adding chaotropic salt;
- and then
- (G) dialyzing and concentrating the DNA.
6 more elements
Dependencies are inferred from the claim text.