Methods and kits for isolating nucleic acid from an organism
The invention generally relates to methods and kits for isolating nucleic acids from an organism. In certain embodiments, methods of the invention involve contacting a plurality of lytic enzymes to an organism, thereby lysing a cell wall of the organism to release the nucleic acid, and introducing at least one agent to separate the nucleic acid from the lysed cells, thereby isolating the nucleic acid.
1. A method for isolating nucleic acid from an organism, the method comprising:
lysing a cell wall of an organism using a composition comprising a plurality of lytic enzymes and that does not contain non-protein lysing chemicals, thereby releasing nucleic acid from the organism;
introducing at least one agent to separate the nucleic acid from the lysed organism, thereby isolating the nucleic acid; and
introducing at least one liquid biocompatible polymer to the isolated nucleic acid, wherein the biocompatible polymer has a viscosity that allows for the isolated nucleic acids to be used in fluid flow applications, with the proviso that the polymer is not agarose.
2. The method according to claim 1 , wherein prior to introducing the agent, the method further comprises introducing proteinase K.
3. The method according to claim 1 , wherein the agent is a cationic detergent.
4. The method according to claim 1 , wherein the agent is an affinity bead.
5. The method according to claim 4 , wherein the affinity bead is a magnetic bead.
6. The method according to claim 1 , wherein prior to the contacting step, the method further comprises separating the organism from a sample.
7. The method according to claim 6 , wherein the sample is selected from the group consisting of a human tissue or body fluid, an environmental sample, and a food sample.
8. The method according to claim 1 , wherein the plurality of lytic enzymes are selected from the group consisting of achromopeptidase, lysozyme, lysostaphin, mutanolysin, and a combination thereof.
9. The method according to claim 1 , wherein the organism is an unknown organism.
10. The method according to claim 1 , wherein lysis occurs in solution.
11. A method for generating a physical map of a genome of an organism, the method comprising:
contacting an organism with a composition comprising a plurality of lytic enzymes but no non-protein lysing chemicals in order to lyse a cell wall of the organism, thereby releasing nucleic acid from the organism;
introducing at least one agent to separate the nucleic acid from the lysed organism, thereby isolating the nucleic acid;
introducing at least one liquid biocompatible polymer to the isolated nucleic acid, wherein the biocompatible polymer has a viscosity that allows for the isolated nucleic acids to be used in fluid flow applications, with the proviso that the polymer is not agarose;
depositing the nucleic acid onto a substrate such that the nucleic acid is elongated and fixed on the substrate so that the nucleic acid remains accessible for enzymatic reactions;
digesting the nucleic acid enzymatically to produce one or more restriction digests;
imaging the restriction digests; and
constructing an optical map from the restriction digests.
12. The method according to claim 11 , wherein prior to introducing the agent, the method further comprises introducing proteinase K.
13. The method according to claim 11 , wherein prior to the contacting step, the method further comprises separating the organism from a sample.
14. The method according to claim 11 , wherein lysis occurs in solution.
15. The method according to claim 11 , wherein the agent is a cationic detergent.
16. The method according to claim 11 , wherein the agent is an affinity bead.