Methods and kits for extraction of DNA
Methods and materials are disclosed for use in recovering a biopolymer from a solution. In particular, the invention provides methods for extraction and isolation of nucleic acids from biological materials. The nucleic acids can be separated by forming a stable complex with soluble polysaccharide polymers and magnetic particles, in the presence of detergents and solvent. When the particles are magnetically separated out of the solution, the nucleic acids are separated with them. The nucleic acids can subsequently be released and separated from the particles. The nucleic acid preparation is useful for achieving efficient and accurate results in downstream molecular techniques such as quantification, identification of the source of the nucleic acids, and genotyping.
1. A method of making a product, wherein the product comprises a nucleic acid, the method comprising the steps of:
(a) treating a sample containing nucleic acids with a starting solution comprising a soluble polymer and detergent to form a first complex wherein the nucleic acids are entrapped by the soluble polymer;
(b) applying suspended magnetically attractable particles to the sample to form a second complex wherein the soluble polymer is attached to the magnetically attractable particles, thereby indirectly connecting the nucleic acids to the magnetically attractable particles; and
(c) recovering the nucleic acids from the sample by applying a magnetic field.
2. The method of claim 1 , wherein the nucleic acids are derived from a sample of biological material selected from the group consisting of blood, blood stain, saliva, saliva stain, buccal cells, buccal swab, semen, semen stain, cigarette butt, chewing gum and combinations thereof.
3. The method of claim 1 , wherein the magnetically attractable particles comprise dextran-encased magnetite nanoparticles.
4. The method of claim 1 , wherein the soluble polymer is selected from the group consisting of polyhydroxy polysaccharides of dextran, cellulose, cellulose derivatives, soluble starch, dextrin, cellodextrin, polyethylene glycol, heparin, glycogen, and combinations thereof.
5. The method of claim 1 , wherein the detergent is selected from the group consisting of N-lauroyl sarcosine, sarcosyl, deoxycholate, CTAB, deoxycholate, dodecyl ß-D-maltoside, nonanoyl-N-methylglucamide, polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether, sodium dodecyl sulfate, and combinations thereof.
6. A method of making a product, wherein the product comprises a nucleic acid, comprising the steps of:
(a) lysing cells from a sample in a lysis solution, thus forming a lysate comprising nucleic acids;
(b) treating the lysate with a starting solution comprising a soluble polymer and detergent to form a first complex wherein the nucleic acids are entrapped by the soluble polymer;
(c) applying suspended magnetically attractable particles to the lysate to form a nucleic acid-polymer-particle complex wherein the soluble polymer is attached to the magnetically attractable particles, thereby indirectly connecting the nucleic acids to the magnetically attractable particles;
(d) washing the nucleic acid-polymer-particle complex with a wash solution; and
(e) recovering the nucleic acids by applying a magnetic field.
7. The method of claim 6 , wherein the magnetically attractable particles comprise dextran-encased magnetite nanoparticles.
8. The method of claim 6 , wherein the soluble polymer is selected from the group consisting of polyhydroxy polysaccharides of dextran, cellulose, cellulose derivatives, soluble starch, dextrin, cellodextrin, polyethylene glycol, heparin, glycogen, and combinations thereof.
9. The method of claim 6 , wherein the detergent is selected from the group consisting of N-lauroyl sarcosine, sarcosyl, deoxycholate, CTAB, deoxycholate, dodecyl β-D-maltoside, nonanoyl-N-methylglucamide, polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether, sodium dodecyl sulfate, and combinations thereof.
10. A method of recovering nucleic acids from a sample, comprising the steps of:
(a) treating the sample with a starting solution comprising a soluble polymer and a detergent to form a first complex wherein the nucleic acids are entrapped by the soluble polymer;
(b) adding suspended magnetically attractable particles to the treated sample to form a second complex wherein the soluble polymer is attached to the magnetically attractable particles, thereby indirectly connecting the nucleic acids to the magnetically attractable particles; and
(c) separating the nucleic acids attached to the magnetically attractable particles via the soluble polymer by applying a magnetic field.
11. The method of claim 10 , further comprising the step of releasing the nucleic acids from the magnetically attractable particles.
12. The method of claim 11 , further comprising the step of eluting the nucleic acids in an aqueous solution.
13. The method of claim 10 , wherein the nucleic acids are derived from a sample of biological material selected from the group consisting of blood, blood stain, saliva, saliva stain, buccal cells, buccal swab, semen, semen stain, cigarette butt, chewing gum, and combinations thereof.
14. The method of claim 10 , wherein the magnetically attractable particles comprise dextran-encased magnetite nanoparticles.
15. The method of claim 10 , wherein the soluble polymer is selected from the group consisting of polyhydroxy polysaccharides of dextran, cellulose, cellulose derivatives, soluble starch, dextrin, cellodextrin, polyethylene glycol, heparin, glycogen, and combinations thereof.
16. The method of claim 10 , wherein the detergent is selected from the group consisting of N-lauroyl sarcosine, sarcosyl, deoxycholate, CTAB, deoxycholate, dodecyl β-D-maltoside, nonanoyl-N-methylglucamide, polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether, sodium dodecyl sulfate, and combinations thereof.