# US 10,246,735 B2 · Methods for preparing samples for nucleic acid amplification > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,246,735 B2 - Document ID: US10246735B2 - Status: Granted - Kind: B2 - Application number: 14/389,526 - Filed: 2013-03-28 - Published: 2015-03-19 - Granted: 2019-04-02 - Earliest claimed: 2012-03-30 - Assignees: Roche Molecular Systems, Inc. - Applicants: Lumora Ltd. - Inventors: Clint Pereira; Cathal Joseph McElgunn; Laurence Carlo Tisi - Web page: https://patentgrep.com/patent/US10246735B2 ## CPC - C12Q 1/6806: Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) - B01D 15/161: {Temperature conditioning} - B01D 15/3876: {modifying the temperature} - B01L 3/502: {with fluid transport, e.g. in multi-compartment structures} - C12N 15/1006: {by means of a solid support carrier, e.g. particles, polymers} - C12N 15/101: {by chromatography, e.g. electrophoresis, ion-exchange, reverse phase} - B01L 2300/0681: Filter - B01L 2300/0832: cylindrical, tube shaped - B01L 2400/0442: thermal energy, e.g. vaporisation, bubble jet - 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 - B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28) - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present invention is in the field of sample preparation. In particular, it relates to methods for preparing samples prior to performing nucleic acid amplification. ## Claims ### Claim 1 A method for passing a liquid sample through a porous solid matrix, comprising the steps of sealing the liquid sample within a container which comprises a porous solid matrix as at least a part of the container and raising a temperature to increase pressure inside the container, thereby to cause the liquid sample to pass through the porous solid matrix, wherein the liquid sample includes a biological sample and a buffer. ### Claim 2 (depends on 1) The method of claim 1, wherein the container comprises two or more different solid porous matrices. ### Claim 3 (depends on 1) The method of claim 1, wherein the liquid sample comprises nucleic acids and inhibitors of nucleic acid amplification. ### Claim 4 (depends on 3) The method of claim 3, wherein the container comprises a porous solid matrix which binds nucleic acids more strongly than inhibitors of nucleic acid amplification. ### Claim 5 (depends on 3) The method of claim 3, wherein the container comprises a porous solid matrix which binds inhibitors of nucleic acid amplification more strongly than nucleic acids. ### Claim 6 A method for purifying nucleic acids from a liquid sample which comprises nucleic acids and inhibitors of nucleic acid amplification, wherein the method comprises the steps of (a) contacting the liquid sample with a porous solid matrix which binds inhibitors of nucleic acid amplification more strongly than nucleic acids, wherein heat is applied to the porous solid matrix and the liquid sample; and (b) separating the liquid sample comprising unbound nucleic acids from the porous solid matrix, wherein the liquid sample is passed through the porous solid matrix by sealing the liquid sample within a container comprising the porous solid matrix and the heat that is applied raises a temperature to increase pressure inside the container, thereby to cause the liquid to pass through the porous solid matrix, and the liquid sample includes a buffer. ### Claim 7 (depends on 6) The method of claim 6, wherein heat is applied to the liquid sample in step (b). ### Claim 8 (depends on 1) The method of claim 1, wherein the container comprises a flow restrictor which is configured to reduce flow of liquid sample from the container through the porous solid matrix compared to a container which does not have the flow restrictor. ### Claim 9 (depends on 8) The method of claim 8, wherein the flow restrictor is a filter, a frit, or a valve. ### Claim 10 (depends on 8) The method of claim 8, wherein the flow restrictor is a layer of material which melts at a temperature between 45° C. and 110° C. ### Claim 11 (depends on 10) The method of claim 10 wherein the layer of material is a wax. ### Claim 12 (depends on 1) The method of claim 1, further comprising a step of lysing the sample. ### Claim 13 (depends on 3) An apparatus for purifying nucleic acids according to claim 3, comprising a) the container comprising the porous solid matrix as at least a part of the container and means for sealing the container, the matrix being capable of binding inhibitors of nucleic acid amplification more strongly than nucleic acids, and b) a heating element configured to heat the container to a temperature of up to 110° C.; wherein the apparatus is configured to pass liquid through the porous solid matrix by heat. ### Claim 14 (depends on 13) The apparatus of claim 13, wherein the apparatus further comprises a second container to receive liquid passed through the porous solid matrix. ### Claim 15 (depends on 13) The apparatus of claim 13, wherein the apparatus further comprises a vessel comprising reagents for nucleic acid amplification. ### Claim 16 (depends on 6) An apparatus for purifying nucleic acids according to claim 6, comprising a) a container comprising a porous solid matrix as at least a part of the container and means for sealing the container, the matrix being capable of binding inhibitors of nucleic acid amplification more strongly than nucleic acids, and b) a heating element configured to heat the container to a temperature of up to 110° C.; wherein the apparatus is configured to pass liquid through the porous solid matrix by heat. ### Claim 17 (depends on 16) The apparatus of claim 16, wherein the apparatus further comprises a second container to receive liquid passed through the porous solid matrix. ### Claim 18 (depends on 16) The apparatus of claim 16, wherein the apparatus further comprises a vessel comprising reagents for nucleic acid amplification.