Fabrication of hierarchical silica nanomembranes and uses thereof for solid phase extraction of nucleic acids

Patent metadata

Field	Value
Publication number	US-10023859-B2
Application number	US-201615382807-A
Country	US
Kind code	B2
Filing date	Dec 19, 2016
Priority date	Aug 5, 2013
Publication date	Jul 17, 2018
Grant date	Jul 17, 2018

Abstract

The present invention provides a novel method to fabricate silica nanostructures on thin polymer films based on silica deposition and self-wrinkling induced by thermal shrinkage. These micro- and nano-scale structures have vastly enlarged the specific area of silica, thus the silica nanomembranes can be used for solid phase extraction of nucleic acids. The inventive silica nanomembranes are suitable for nucleic acid purification and isolation and demonstrated better performance than commercial particles in terms of DNA recovery yield and integrity. In addition, the silica nanomembranes have extremely high nucleic acid capacity due to its significantly enlarged specific surface area of silica. Methods of use and devices comprising the silica nanomembranes are also provided.

First claim

The invention claimed is:

1. A method for extracting nucleic acids from a sample comprising:
a) contacting the sample comprising nucleic acids with a sufficient amount of silica nanomembranes, wherein the silica nanomembranes comprise a heat shrunken polymer core coated with a silicon dioxide layer, and wherein the silicon dioxide layer comprises a surface structure comprising wrinkle structures having a length from about 10 nm to about 100 μm and/or discreet substantially planar flakes having a length from about 10 nm to about 100 μm; and
b) purifying said nucleic acids from the sample.

2. The method of claim 1 , wherein the sample is selected from the group consisting of: a cell, tissue, blood, serum body fluid, formalin-fixed paraffin embedded tissue, enzymatic reaction, plant, bacteria, and viral particle.

3. The method of claim 1 , further comprising at step a) deparaffinizing the sample.

4. The method of claim 3 , wherein washing comprises two or more washes.

5. The method of claim 3 , further comprising a drying step after washing.

6. The method of claim 3 , wherein deparaffinization comprises contacting the sample with an organic solvent.

7. The method of claim 6 , wherein the organic solvent is selected from the group consisting of: xylene, mineral oil, hexadecane, toluene, 5-chloro-2-methyl-4-isothiazolin-3-one, 5-chloro-2-methyl-4-isothiazolin-3-one; a terpene or isoparaffinic hydrocarbon, and 2-butoxyethanol.

8. The method of claim 6 , wherein step a) further comprises removing the organic solvent, and washing the sample.

9. The method of claim 1 , further comprising step c) desorbing the nucleic acids from the silica nanomembranes to obtain the isolated and purified nucleic acids from the sample.

10. The method of claim 9 , wherein at step c) the desorption of the nucleic acids from the silica nanomembranes is by application of an elution solution.

11. The method of claim 1 , wherein purifying said nucleic acids comprises washing the silica nanomembranes to remove any non-desired components.

12. The method of claim 1 , further comprising contacting the sample with a lysis and/or digestion solution prior to step a).

13. The method of claim 1 , wherein at step a) the nucleic acids are contacted with a chaotropic agent.

14. The method of claim 1 , further comprising at step a) contacting the sample with the sufficient amount of silica nanomembranes in the presence of an aqueous alcoholic solution.

15. The method of claim 1 , further comprising adding to the sample a DNAse or an RNAse to allow removal of non-desired DNA or RNA components prior to step a).

16. The method of claim 1 , wherein the nucleic acids are DNA.

17. The method of claim 1 , wherein the nucleic acids are RNA.

18. The method of claim 1 , wherein the nucleic acids are selected from the group consisting of: plasmid, genomic, mitochondrial, vesicle, and cell free sources.

19. The method of claim 1 , wherein the nucleic acids bind to the silica nanomembranes.

Assignees

• Univ Johns Hopkins

Inventors

• Wang Tza-Huei
• Zhang Yi

Classifications

• B01D69/02

  characterised by their properties · CPC title

• B01D2325/06

  Surface irregularities · CPC title

• B01D63/087

  Single membrane modules · CPC title

• B01J20/3234

  Inorganic material layers · CPC title

• C12N15/1006 Primary

  by means of a solid support carrier, e.g. particles, polymers · CPC title