Method of isolating nucleic acid
View Patent ↗Disclosed herein is a method for isolating a nucleic acid from a sample. The method includes contacting the sample with boron carbide under conditions sufficient to form a boron carbide-nucleic acid complex. The complex is separated from the sample.
1. A method for isolating nucleic acids from a sample, the method comprising:
(a) contacting the sample with a boron carbide composition under conditions sufficient to form a boron carbide-nucleic acid complex;
(b) separating the complex from the sample; and
(c) eluting the nucleic acids from the complex, thereby isolating the nucleic acids.
2. The method of claim 1 , wherein the sample is prepared in the absence of an organic extraction.
3. The method of claim 2 , wherein the organic extraction comprises phenol and chloroform.
4. The method of claim 3 , wherein the phenol is an acid phenol.
5. The method of claim 1 , further comprising contacting the sample with a chaotrope.
6. The method of claim 5 , wherein the chaotrope is present in a lysis or binding buffer.
7. The method of claim 5 , wherein the chaotrope is guanidine thiocyanate (GTC) or guanidine hydrochloride (GHCl).
8. The method of claim 1 , wherein the boron carbide composition further comprises glass fibers.
9. The method of claim 1 , wherein the boron carbide-nucleic acid complex is formed in the presence of an alcohol.
10. The method of claim 9 , wherein the alcohol is isopropanol or ethanol.
11. The method of claim 1 , wherein formation of the boron carbide-nucleic acid complex comprises selective binding of the nucleic acid to the boron carbide and wherein selective binding is dependent upon a concentration of an alcohol.
12. The method of claim 11 , wherein the alcohol is isopropanol or ethanol.
13. The method of claim 12 , wherein the nucleic acid is RNA and wherein the RNA selectively binds the boron carbide when the concentration of alcohol is from about 15% v/v to about 100% v/v.
14. The method of claim 13 , wherein the RNA is large RNA and wherein the large RNA selectively binds the boron carbide when the concentration of isopropanol is from about 15% to about 20%.
15. The method of claim 13 , wherein the RNA is large RNA and wherein the large RNA selectively binds the boron carbide when the concentration of ethanol is from about 20% v/v to about 35% v/v.
16. The method of claim 13 , wherein the RNA is small RNA and wherein the small RNA selectively binds the boron carbide when the concentration of isopropanol is about 40% v/v to about 60% v/v.
17. The method of claim 13 , wherein the RNA is small RNA and wherein the small RNA selectively binds the boron carbide when the concentration of ethanol is about 50% v/v to about 70% v/v.
18. The method of claim 14 , wherein the RNA is long, noncoding RNA.
19. The method of claim 15 , wherein the small RNA is selected from the group consisting of microRNA (miRNA), small interfering RNA (siRNA), piwi-interacting RNA (piRNA), small noncoding RNA, small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), and transfer RNA (tRNA).
20. The method of claim 19 , wherein the small RNA is miRNA.
21. The method of claim 1 , wherein the nucleic acid is RNA.
22. The method of claim 21 , wherein the RNA is small RNA.
23. The method of claim 21 , wherein the RNA is microRNA (miRNA).
24. The method of claim 21 , wherein the RNA comprises RNA molecules having a length of about 5 nucleotides to about 200 nucleotides.
25. The method of claim 24 , wherein the RNA comprises RNA molecules having a length of about 10 nucleotides to about 100 nucleotides.
26. The method of claim 24 , wherein the RNA comprises RNA molecules having a length of about 18 nucleotides to about 30 nucleotides.
27. The method of claim 1 , wherein the boron carbide has a particle size of about 1 micron to about 20 microns.
28. The method of claim 27 , wherein the boron carbide particles are in a suspension.
29. The method of claim 1 , wherein the boron carbide is associated with a solid support.
30. The method of claim 29 , wherein the solid support is selected from the group consisting of a magnetic particle and a column.
31. The method of claim 29 , wherein the solid support is used in a magnetic, centrifugation, or filtration format.
32. The method of claim 1 , further comprising using or characterizing the isolated nucleic acid.