# US 8,771,955 B2 · Device and method for extraction and analysis of nucleic acids from biological samples

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- Number: US 8,771,955 B2
- Document ID: US8771955B2
- Status: Granted
- Kind: B2
- Application number: 13/435,440
- Filed: 2012-03-30
- Published: 2012-10-04
- Granted: 2014-07-08
- Earliest claimed: 2006-06-26
- Assignees: Blood Cell Storage, Inc.
- Applicants: Michael W. Reed; Oliver Z. Nanassy
- Inventors: Michael W. Reed; Oliver Z. Nanassy
- Web page: https://patentgrep.com/patent/US8771955B2

## CPC

- C12Q 1/6851: Quantitative amplification
- B01L 3/5027: {by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip}
- B01L 3/50273: {characterised by the means or forces applied to move the fluids}
- C12Q 1/6806: Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence)
- B01L 7/525: {with physical movement of samples between temperature zones}
- B01L 2200/10: Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
- B01L 2300/0816: Cards, e.g. flat sample carriers usually with flow in two horizontal directions
- B01L 2300/0887: Laminated structure
- B01L 2400/0457: passive flow or gravitation
- B01L 2400/0487: fluid pressure, pneumatics
- C12Q 1/686: Polymerase chain reaction [PCR]
- Y10T 436/143333: Saccharide [e.g., DNA, etc.]
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

Device and methods for extracting and analyzing nucleic acids from biological samples.

## Claims

### Claim 1

A method for quantifying and amplifying the amount of nucleic acids in a sample, comprising:
(a) contacting nucleic acids from a sample with a nucleic acid amplification reaction mixture comprising a first fluorescent compound having a first fluorescence emission maximum, a second fluorescent compound having a second fluorescence emission maximum, and a third fluorescent compound having a third fluorescence emission maximum and a fluorescence intensity dependent on the concentration of amplified nucleic acids, wherein the first, second, and third fluorescence emission maxima are different, each having a fluorescence intensity dependent on the concentration of nucleic acids;
(b) measuring the fluorescence of the first fluorescent compound to determine the amount of nucleic acids;
(c) treating the nucleic acids and the nucleic acid amplification reaction mixture under conditions for amplifying the nucleic acids to provide amplified nucleic acids; and
(d) measuring the fluorescence of the first fluorescent compound to determine the amount of amplified nucleic acids.

### Claim 2 (depends on 1)

The method of claim 1, wherein the first fluorescent compound is a minor groove binder or an intercalating agent.

### Claim 3 (depends on 1)

The method of claim 1 further comprising measuring the fluorescence of the second fluorescent compound to determine the presence of a gene or gene product.

### Claim 4 (depends on 1)

The method of claim 1, wherein the second fluorescent compound is an oligonucleotide probe.

### Claim 5 (depends on 1)

The method of claim 1 further comprising measuring the fluorescence of the third fluorescent compound to determine the presence of a gene or gene product.

### Claim 6 (depends on 1)

The method of claim 1 wherein the third fluorescent compound is an oligonucleotide probe.

### Claim 7

A method for quantifying and amplifying the amount of nucleic acids in a sample, comprising:
(a) contacting nucleic acids from a sample with a first fluorescent compound having a first fluorescence emission maximum and a fluorescence intensity dependent on the concentration of nucleic acids;
(b) measuring the fluorescence of the first fluorescent compound to determine the amount of nucleic acids;
(c) treating the nucleic acids with a nucleic acid amplification reaction mixture under conditions for amplifying the nucleic acids to provide amplified nucleic acids, wherein the nucleic acid amplification reaction mixture comprises a second fluorescent compound having a second fluorescence emission maximum and a fluorescence intensity dependent on the concentration of nucleic acids, and a third fluorescent compound having a third fluorescence emission maximum and a fluorescence intensity dependent on the concentration of nucleic acids, wherein the first, second, and third fluorescence emission maxima are different; and
(d) measuring the fluorescence of the second fluorescent compound to determine the presence of a gene or gene product.

### Claim 8 (depends on 7)

The method of claim 7 further comprising measuring the fluorescence of the first fluorescent compound to determine the amount of amplified nucleic acids.

### Claim 9 (depends on 7)

The method of claim 7, wherein the first fluorescent compound is a minor groove binder or an intercalating agent.

### Claim 10 (depends on 7)

The method of claim 7, wherein the second fluorescent compound is an oligonucleotide probe.

### Claim 11 (depends on 7)

The method of claim 7, wherein the third fluorescent compound is an oligonucleotide probe.

### Claim 12 (depends on 7)

The method of claim 7 further comprising measuring the fluorescence of the third fluorescent compound to determine the presence of a gene or gene product.