LYSIS BUFFERS FOR EXTRACTING NUCLEIC ACIDS
The present teachings relate to the extraction of nucleic acid from solid materials. Provided are useful compositions, methods and kits for obtaining nucleic acids from a solid biological sample or an adhesive material having a biological material adherent or embedded within the adhesive substrate. The extracted nucleic acid can be used in downstream applications such as genotyping, detection, quantification, and identification of the source of the biological material.
1 .- 37 . (canceled)
38 . A method of making a nucleic acid, comprising:
i. lysing a solid sample in the lysis solution of the kit of claim 76 , forming a lysate comprising the nucleic acid;
ii. extracting the lysate product, wherein the nucleic acid is in the lysate.
39 . The method of claim 38 , wherein the solid sample comprises the nucleic acid.
40 . The method of claim 39 , wherein the solid is a biological material, an adhesive material or substrate, or a natural or synthetic substrate.
41 . The method of claim 40 , wherein the biological material is selected from bone, cartilage, ligament, tendon, and tooth.
42 . The method of claim 40 , wherein the adhesive material substrate comprises a biological sample.
43 . The method of claim 42 , wherein the biological sample is embedded in the adhesive material substrate.
44 . The method of claim 43 , wherein the adhesive material is selected from chewing gum, cigar butt, cigarette butt, adhesive film, adhesive label, adhesive paper, adhesive skin patch, envelope, stamp, a fingerprint tape lift, and adhesive tape.
45 . The method of claim 44 , wherein the adhesive skin patch is selected from the group consisting of electronic electrode, transferable tattoo, transdermal chemical substance patch and would care dressing.
46 .- 68 . (canceled)
69 . The method according to claim 38 , wherein the solid sample is a fingerprint tape lift.
70 .- 75 . (canceled)
76 . A kit for extracting nucleic acid from a solid sample comprising:
a lysis buffer comprising 250 to 500 mM ethylene glycol tetraacetic acid, a detergent, a reducing agent, and an enzyme,
wherein the detergent is selected from one or more of N-lauroyl sarcosine, sodium deoxycholate, cetyltrimethylammonium bromide, dodecyl β-D-maltoside, nonanoyl-N-methylglucamide, polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether, sodium dodecyl sulfate, and combinations thereof, wherein the reducing agent comprises one or more of tris(2-carboxyethyl)phosphine, dithioerythritol, and dithiothreitol, and
wherein the enzyme is selected from caspase, chymotrypsin, pepsin, proteinase K, thrombin, Staphylococcus V8 protease, pronase, papain, Bacillus sp. E1A protease, and trypsin and combinations thereof.
77 .- 81 . (canceled)
82 . The kit of claim 76 , wherein the solid sample is a biological material or an adhesive material.
83 . The kit of claim 82 , wherein the biological material is selected from bone, cartilage, ligament, tendon, and tooth.
84 . The kit of claim 83 , wherein the adhesive material is selected from chewing gum, cigar butt, cigarette butt, adhesive film, adhesive label, adhesive paper, adhesive skin patch, envelope, stamp, a fingerprint tape lift, and adhesive tape.
85 . The kit of claim 84 , wherein the adhesive skin patch is selected from the group consisting of electronic electrode, transferable tattoo, transdermal chemical substance patch and would care dressing.
86 . The kit of claim 76 , further comprising:
a solution comprising a polymer and detergent; and
magnetically attractable particles.
87 . The kit of claim 86 , wherein the polymer comprises one or more of dextran, cellulose, cellulose derivatives, soluble starch, dextrin, cellodextrin, polyethylene glycol, heparin, glycogen, and combinations thereof and
wherein the detergent comprises one or more of N-lauroyl sarcosine, sodium deoxycholate, CTAB, dodecyl β-D-maltoside, nonanoyl-N-methylglucamide, polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether, sodium dodecyl sulfate, and combinations thereof.
88 . (canceled)
89 . The kit of claim 86 , wherein the magnetically attractable particles comprise dextran-encased magnetic nanoparticles.
90 . (canceled)
91 . The kit of claim 86 , further comprising a magnetic device.
92 . The kit of claim 86 , further comprising a wash solution.
93 . The kit of claim 1 , further comprising a salt selected from KCl, NaCl, MgCl, MnCl 2 .
94 .- 100 . (canceled)