Compositions and methods for nucleic acid extraction

Patent metadata

Field	Value
Publication number	US-9410147-B2
Application number	US-201414212799-A
Country	US
Kind code	B2
Filing date	Mar 14, 2014
Priority date	Mar 15, 2013
Publication date	Aug 9, 2016
Grant date	Aug 9, 2016

Abstract

Compositions and techniques for the extraction, enrichment and isolation of nucleic acids from fresh, fixed or fixed and embedded cells, tissues, biological materials and cellular source material using amine monomers are disclosed herein.

First claim

What is claimed is:

1. A method of isolating an extraction solution comprising nucleic acids from cellular source material, said method comprising:
a) providing i) cellular source material and ii) an aqueous extraction solution comprising 3-amino-1-propanol, a chaotrope and a detergent;
b) contacting said cellular source material with said extraction solution resulting in lysis of the cellular material and the isolation of an extraction solution comprising nucleic acids.

2. The method of claim 1 , wherein said chaotrope is selected from the group consisting of urea, guanidine thiocyanate (GITC), ethanol and butanol.

3. The method of claim 1 , wherein said detergent is selected from the group consisting of polysorbates, deoxycholate, sodium deoxycholate and sodium dodecyl sulfate (SDS), nonyl phenoxypolyethoxylethanol and polyethylene glycol.

4. The method of claim 3 , wherein the concentration of said detergent is about 8% to about 15%.

5. The method of claim 1 , further comprising wherein said alcohol is selected from the group consisting of ethanol and butanol.

6. The method of claim 5 , wherein the concentration of said alcohol is about 15% to about 25%.

7. The method of claim 1 , wherein the concentration of 3-amino-1-propanol in said aqueous extraction solution is about 30% to about 50%.

8. The method of claim 1 , wherein the concentration of the chaotrope in said aqueous extraction solution is about 4 M to about 5 M.

9. The method of claim 1 , wherein said cellular source material is selected from living cellular source material and fixed cellular source material.

10. The method of claim 9 , wherein said living cellular source material comprises a suspension of single cells.

11. The method of claim 10 , wherein said suspension of cells comprise bacteria.

12. The method of claim 11 , wherein said bacteria are Mycobacteria.

13. The method of claim 9 , wherein said suspension of cells comprise yeast.

14. The method of claim 9 , wherein said fixed cellular source material comprises formalin-fixed paraffin embedded (FFPE) material.

15. The method of claim 1 , further comprising wherein said aqueous extraction solution is enzyme-free.

16. The method of claim 1 , further comprising wherein said aqueous extraction solution is protease-free.

17. A composition comprising an aqueous extraction solution suitable for the extraction of nucleic acids from cellular source material, said composition comprising 3-amino-1-propanol, a chaotropic reagent, a detergent and an organic solvent.

18. The composition of claim 17 , wherein said chaotrope is selected from the group consisting of urea, guanidine thiocyanate (GITC), ethanol and butanol.

19. The composition of claim 18 , wherein the concentration of the chaotrope in said aqueous extraction solution is about 4 M to about 5 M.

20. The composition of claim 17 , wherein said detergent is selected from the group consisting of polysorbates, deoxycholate, sodium deoxycholate and sodium dodecyl sulfate (SDS), nonyl phenoxypolyethoxylethanol and polyethylene glycol.

21. The composition of claim 17 , wherein the concentration of said detergent is about 8% to about 15%.

22. The composition of claim 17 , wherein said organic solvent is an alcohol and said alcohol is selected from the group consisting of ethanol and butanol.

23. The composition of claim 22 , wherein the concentration of said alcohol is about 15% to about 25%.

24. The composition of claim 17 , wherein the concentration of 3-amino-1-propanol in said aqueous extraction solution is about 30% to about 50%.

Assignees

• Abbott Molecular Inc

Inventors

• Gundling Gerard J

Classifications

• C12N15/1013

  by using magnetic beads · CPC title

• C12N15/1003 Primary

  Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor · CPC title

• C07H1/08

  from natural products · CPC title

• C12Q1/6806

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• C07H19/16

  Purine radicals · CPC title