Title: Method and apparatus for purifying nucleic acids and performing polymerase chain reaction assays using an immiscible fluid

Abstract

Embodiments of the present invention are directed toward devices, systems, and methods for purifying nucleic acids to conduct polymerase chain reaction (PCR) assays. In one example, a method includes generating complexes of silica beads and nucleic acids in a lysis buffer, transporting the complexes through an immiscible fluid to remove interfering compounds from the complexes, further transporting the complexes into a density medium containing components required for PCR where the nucleic acids disassociate from the silica beads, and thermocycling the contents of the density medium to achieve PCR. Signal may be detected from labeling agents in the components required for PCR.

Inventors:

Koh, Chung-Yan; Light, Yooli Kim; Piccini, Matthew Ernest; Singh, Anup K.

Issue Date:

2017-10-31

Research Org.:

Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

Sponsoring Org.:

USDOE

OSTI Identifier:

1406570

Patent Number(s):

9803238

Application Number:

14/090,040

Assignee:

National Technology & Engineering Solutions of Sandia, LLC

Patent Classifications (CPCs):

B - PERFORMING OPERATIONS B01 - PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL B01L - CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE

C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS

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B - PERFORMING OPERATIONS B01 - PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL B01L - CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE

B01L2300/087 - Multiple sequential chambers
B01L3/502761 - {specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
B01L2400/0409 - centrifugal forces
B01L2300/0803 - Disc shape
B01L2200/0668 - Trapping microscopic beads
B01L7/52 - {with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
B01L2300/1827 - using resistive heater

C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS

C12Q1/686 - Polymerase chain reaction [PCR]

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DOE Contract Number:  

AC04-94AL85000

Resource Type:

Patent

Resource Relation:

Patent File Date: 2013 Nov 26

Country of Publication:

United States

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES