# US 9,040,675 B2 · Formulations for nucleic acid stabilization on solid substrates

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 9,040,675 B2
- Document ID: US9040675B2
- Status: Granted
- Kind: B2
- Application number: 13/721,948
- Filed: 2012-12-20
- Published: 2013-10-31
- Granted: 2015-05-26
- Earliest claimed: 2012-04-30
- Assignees: General Electric Company
- Applicants: General Electric Company
- Inventors: Brian Christopher Bales; Erik Leeming Kvam; Jason Louis Davis
- Web page: https://patentgrep.com/patent/US9040675B2

## CPC

- B01J 20/045: {containing sulfur, e.g. sulfates, thiosulfates, gypsum}
- B01J 20/24: Naturally occurring macromolecular compounds, e.g. humic acids or their derivatives
- C12N 15/1006: {by means of a solid support carrier, e.g. particles, polymers}
- C12N 15/1017: {by filtration, e.g. using filters, frits, membranes}
- G01N 1/40: Concentrating samples
- B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

The present disclosure generally relates to dry solid matrices for the extraction, stabilization, and storage of nucleic acids, particularly RNA, in a dry format under ambient conditions for a prolonged period of time. Methods for collecting and recovering the nucleic acids stored in the dry solid matrix are also described.

## Claims

### Claim 1

A dry solid matrix for extraction and storage of nucleic acids from a biological sample, wherein a composition comprising;
a metal thiocyanate comprises a Group 1 or Group 2 metal cation;
a reducing agent where the reducing agent is tris(2-carboxyethyl)phosphine (TCEP); and
a buffer; is incorporated into the dry solid matrix, and the matrix is then dried; and where the solid matrix is a porous matrix comprising cellulose, cellulose acetate, glass fiber, or any combination thereof.

### Claim 2 (depends on 1)

The dry solid matrix of claim 1, wherein the Group 1 or Group 2 metal cation is selected from the group consisting of Na+, K+, Li+, Mg+2, Ca+2, and Ba+2.

### Claim 3 (depends on 1)

The dry solid matrix of claim 1, wherein the composition present in the solid matrix further comprises a UV inhibitor, a free-radical trap, a chelator, or any combination thereof

### Claim 4 (depends on 1)

The dry solid matrix of claim 1, wherein the composition incorporated into the dry solid matrix further comprises an RNase inhibitor.

### Claim 5 (depends on 1)

The dry solid matrix of claim 1, wherein the dry solid matrix permits prolonged storage of nucleic acids in a dry format under ambient conditions.

### Claim 6 (depends on 5)

The dry solid matrix of claim 5, wherein the nucleic acids are RNA, DNA, or a combination thereof.

### Claim 7 (depends on 6)

The dry solid matrix of claim 6, wherein the nucleic acids are RNA.

### Claim 8 (depends on 1)

The dry solid matrix of claim 1, wherein the porous matrix is a cellulose paper.

### Claim 9 (depends on 1)

The dry solid matrix of claim 1, wherein the metal thiocyanate is selected from the group consisting of sodium thiocyanate, potassium thiocyanate, lithium thiocyanate, magnesium thiocyanate, calcium thiocyanate, barium thiocyanate, zinc thiocyanate, and a combination thereof.

### Claim 10 (depends on 1)

The dry solid matrix of claim 1, wherein the buffer is selected from the group consisting of 2-amino-2-hydroxymethyl-propane-1,3-diol (Tris), 2-(N-morpholino)ethanesulfonic acid (MES), 3-(N-morpholino)propanesulfonic acid (MOPS), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), a citrate buffer, and a phosphate buffer.

### Claim 11 (depends on 10)

The dry solid matrix of claim 10, wherein the pH range is between 3 and 6.

### Claim 12 (depends on 3)

The dry solid matrix of claim 3, wherein the UV protectant or free-radical trap is selected from the group consisting of hydroquinone monomethyl ether (MEHQ), hydroquinone (HQ), toluhydroquinone (THQ), and ascorbic acid.

### Claim 13 (depends on 4)

The dry solid matrix of claim 4, wherein the RNase inhibitor is vanadyl ribonucleoside complex (VRC) or a commercially available RNase inhibitor.

### Claim 14 (depends on 13)

The solid matrix of claim 13 further comprising a free-radical trap, wherein the free-radical trap comprises MEHQ or THQ.

### Claim 15

A method for extracting and storing nucleic acids from a sample comprising:
a) providing a dry solid matrix for extraction and storage of nucleic acids from a biological sample, where the solid matrix is a porous matrix comprising cellulose, cellulose acetate, glass fiber, or any combination thereof, and wherein a composition is incorporated into the dry solid matrix comprising,
a metal thiocyanate comprises a Group 1 or Group 2 metal cation;
a reducing agent ,where the reducing agent is TCEP;
and a buffer;
b) drying the solid matrix after incorporation of the composition into the dry solid matrix;
c) applying a sample to the dry solid matrix to collect the nucleic acids;
d) drying the solid matrix comprising the nucleic acids; and
e) storing the nucleic acids on the solid matrix in a dry state under ambient conditions.

### Claim 16 (depends on 15)

The method of claim 15, wherein the metal cation is selected from the group consisting of Na+, K+, Li+, Mg+2, Ca+2, and Ba+2.

### Claim 17 (depends on 15)

The method of claim 15, wherein the method further comprises recovering the nucleic acids from the solid matrix.

### Claim 18 (depends on 15)

The method of claim 15, wherein the biological sample is blood, serum, tissue, saliva, nasal mucous, or cells.

### Claim 19 (depends on 15)

The method of claim 15, wherein the sample is a purified nucleic acid sample or a tissue culture cell preparation.

### Claim 20 (depends on 15)

The method of claim 15, wherein purification of the nucleic acids from the sample is not required prior to applying the sample to the dry solid matrix for extraction and storage of the nucleic acids.

### Claim 21 (depends on 15)

The method of claim 15, wherein the method permits the prolonged storage of RNA in a dry format under ambient conditions.

### Claim 22 (depends on 17)

The method of claim 17, wherein the nucleic acids are recoved from the dry solid matrix by rehydrating the matrix in an aqueous solution, a buffer, or an organic solution, and wherein the nucleic acids are subjected further analysis.

### Claim 23 (depends on 17)

The method of claim 17, wherein the nucleic acids are recovered from the dry solid matrix by electroelution.