Methods and kits for isolating nucleic acid from an organism

Abstract

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.

What is claimed is: 
     
         1 . A method for isolating nucleic acid from an organism, the method comprising:
 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.   
     
     
         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 , further comprising introducing at least one 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. 
     
     
         4 . The method according to  claim 1 , wherein the agent is a cationic detergent. 
     
     
         5 . The method according to  claim 1 , wherein the agent is an affinity bead. 
     
     
         6 . The method according to  claim 5 , wherein the affinity bead is a magnetic bead. 
     
     
         7 . The method according to  claim 1 , wherein prior to the contacting step, the method further comprises separating the organism from a sample. 
     
     
         8 . The method according to  claim 7 , wherein the sample is selected from the group consisting of a human tissue or body fluid, an environmental sample, and a food sample. 
     
     
         9 . 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. 
     
     
         10 . The method according to  claim 1 , wherein the organism is an unknown organism. 
     
     
         11 . The method according to  claim 1 , wherein lysis occurs in solution. 
     
     
         12 . A kit comprising:
 a plurality of lytic enzymes; and   at least one separating agent.   
     
     
         13 . The kit according to  claim 12 , further comprising proteinase K. 
     
     
         14 . The kit according to  claim 12 , further comprising at least one biocompatible polymer, wherein the polymer has a viscosity that allows for use of nucleic acids in fluid flow applications. 
     
     
         15 . The kit according to  claim 14 , wherein the polymer is selected from the group consisting of methyl cellulose, polyvinylpryollidone (PVP), Ficol, and a combination thereof. 
     
     
         16 . The kit according to  claim 12 , wherein the plurality of lytic enzymes are selected from the group consisting of achromopeptidase, lysozyme, lysostaphin, mutanolysin, and a combination thereof. 
     
     
         17 . The kit according to  claim 12 , wherein the agent is a cationic detergent. 
     
     
         18 . The method according to  claim 12 , wherein the agent is an affinity bead. 
     
     
         19 . The method according to  claim 18 , wherein the affinity bead is a magnetic bead. 
     
     
         20 . A method for generating a physical map of a genome of an organism, the method comprising:
 contacting a plurality of lytic enzymes to an organism, thereby lysing a cell wall of the organism to release the nucleic acid;   introducing at least one agent to separate the nucleic acid from the lysed cells, thereby isolating the nucleic acid;   introducing at least one 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;   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.   
     
     
         21 . The method according to  claim 20 , wherein prior to introducing the agent, the method further comprises introducing proteinase K. 
     
     
         22 . The method according to  claim 20 , wherein prior to the contacting step, the method further comprises separating the organism from a sample. 
     
     
         23 . The method according to  claim 20 , wherein lysis occurs in solution. 
     
     
         24 . The method according to  claim 20 , wherein the agent is a cationic detergent. 
     
     
         25 . The method according to  claim 20 , wherein the agent is an affinity bead.