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. A method of making a product, wherein the product comprises a nucleic acid, comprising:
a) incubating a solid in a lysate solution, the solid comprising an adhesive material associated with a biological material that includes the nucleic acid, wherein the lysate solution comprises a detergent, a chelating agent, a reducing agent, and an enzyme, and wherein
(i) the chelating agent is included at 250-500 mM,
(ii) the reducing agent is included at 10-100 mM, and/or
(iii) the enzyme is included at 0.5-10 mg/ml; and
b) extracting a supernatant, wherein the supernatant comprises the nucleic acid extracted from the solid.
2. The method of claim 1 , wherein the biological material is embedded in the adhesive material.
3. The method of claim 2 , 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.
4. The method of claim 3 , wherein the adhesive skin patch is selected from the group consisting of electronic electrode, transferable tattoo, transdermal chemical substance patch and wound care dressing.
5. The method of claim 1 , wherein the detergent is selected from 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, and sodium dodecyl sulfate.
6. The method of claim 1 , wherein the chelating agent comprises at least one of ethylene glycol tetraacetic acid (EGTA), ethylene diamine tetraacetic acid (EDTA) and citric acid.
7. The method of claim 1 , wherein the reducing agent comprises at least one of tris(2-carboxyethyl)phosphine (TCEP), dithioerythritol (DTE), dithioerythritol (DTE), and dithiothreitol (DTT).
8. The method of claim 1 , wherein the enzyme comprises one or more of caspase, chymotrypsin, pepsin, proteinase K, thrombin, Staphylococcus V8 protease, pronase, papain, Bacillus sp. E1A protease, and trypsin and combinations thereof.
9. The method of claim 1 , further comprising the step of shaking the incubating solution prior to extracting the supernatant product.
10. The method of claim 1 , wherein the supernatant is extracted by centrifuging a mixture comprising the nucleic acid and the lysate solution.
11. A method of separating a nucleic acid from a solid sample, comprising:
(a) lysing the solid sample in a lysis solution, thereby forming a mixture comprising the nucleic acid, wherein the solid sample comprises an adhesive material associated with a biological material that includes the nucleic acid, wherein the lysate solution comprises a detergent, a chelating agent, a reducing agent, and an enzyme, and wherein
(i) the chelating agent is included at 250-500 mM,
(ii) the reducing agent is included at 10-100 mM, and/or
(iii) the enzyme is included at 0.5-10 mg/ml; and
(b) centrifuging the mixture, wherein the nucleic acid is thereby separated from the solid sample.
12. The method of claim 11 , 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.
13. The method of claim 11 ,
wherein the detergent is selected from 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, and sodium dodecyl sulfate,
wherein the chelating agent comprises at least one of ethylene glycol tetraacetic acid (EGTA), ethylene diamine tetraacetic acid (EDTA) and citric acid,
wherein the reducing agent comprises at least one of tris(2-carboxyethyl)phosphine (TCEP), dithioerythritol (DTE), and dithiothreitol (DTT), and
wherein the enzyme comprises one or more of caspase, chymotrypsin, pepsin, proteinase K, thrombin, Staphylococcus V8 protease, pronase, papain, Bacillus sp. E1A protease, and trypsin and combinations thereof.