Rapid methods for the extraction of nucleic acids from biological samples
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12N-015/10
C12Q-001/686
C12Q-001/68
출원번호
US-0972341
(2018-05-07)
등록번호
US-10087439
(2018-10-02)
발명자
/ 주소
Fischer, Gerald W.
Daum, Luke T.
출원인 / 주소
Longhorn Vaccines and Diagnostics, LLC
대리인 / 주소
Remenick PLLC
인용정보
피인용 횟수 :
0인용 특허 :
147
초록▼
The invention is directed to compositions and methods for rapidly and efficiently extracting nucleic acids and/or targeted nucleic acids sequences from biological samples. The methods of the invention comprise combining the sample with a buffer and magnetic silicon beads and concentrating the beads
The invention is directed to compositions and methods for rapidly and efficiently extracting nucleic acids and/or targeted nucleic acids sequences from biological samples. The methods of the invention comprise combining the sample with a buffer and magnetic silicon beads and concentrating the beads with a magnet or other electrical field. Liquid may be removed, or not, and an alkaline buffer is added followed by magnetic carboxy beads in a binding buffer so that nucleic acids transfer to the carboxy beads, which can be easily and quickly isolated once again with a magnet. Total nucleic acid extraction is greatly enhanced. Extracted nucleic acids can be analyzed, for example, by PCR wherein the nucleic acids can be identified and characterized. Carboxy beads may also contain a ligand so as to target specific nucleic acid sequences. The invention is also directed to kits comprising the tools and compositions for performing the methods of the invention.
대표청구항▼
1. A method of extracting nucleic acids from a biological sample containing cells and/or microorganisms comprising: combining the biological sample with a lysis buffer to form a mix;combining the mix with magnetic matrix material to form a solution, wherein neither the buffer or matrix material disr
1. A method of extracting nucleic acids from a biological sample containing cells and/or microorganisms comprising: combining the biological sample with a lysis buffer to form a mix;combining the mix with magnetic matrix material to form a solution, wherein neither the buffer or matrix material disrupt the cells and/or microorganisms of the biological sample and the magnetic matrix material binds to nucleic acids of the biological sample other than those present within the cells and/or microorganisms;exposing the solution to a magnetic field and removing liquid to concentrate the magnetic matrix material;adding an alkaline buffer to the concentrated magnetic matrix material to form a mixture, wherein the alkaline buffer causes the release of nucleic acids from the magnetic matrix material;adding carboxy-modified magnetic beads in a binding buffer to the mixture wherein the carboxy-modified magnetic beads bind to specific nucleic acid sequences;exposing the mixture to a magnetic field and removing liquid to isolate the carboxy-modified magnetic beads bound to the specific nucleic acid sequences; andeluting the specific nucleic acid sequences from the carboxy-modified magnetic beads using purified water and/or a Tris-EDTA buffer. 2. The method of claim 1, wherein the lysis buffer comprises a chaotrope, a detergent, a reducing agent, a buffer, and a chelator at a pH of about 6-8. 3. The method of claim 1, wherein the biological sample combined with buffer is stored for between about 2 days to about 90 days before combining with magnetic matrix material. 4. The method of claim 3, wherein the biological sample combined with buffer is stored for between about 2 days to about 20 days before combining with magnetic matrix material. 5. The method of claim 1, wherein the binding buffer comprises PEG, a salt, a buffering agent, a chelator, a detergent, NLS and an alcohol. 6. The method of claim 1, which does not involve centrifugation. 7. The method of claim 1, which is performed in a single vessel. 8. The method of claim 1, which is automated for high-throughput analysis of a plurality of biological samples. 9. The method of claim 1, further comprising analyzing the specific nucleic acid sequences by a PCR. 10. The method of claim 1, further comprising identifying the specific nucleic acid sequences. 11. The method of claim 10, wherein identifying comprises PCR. 12. The method of claim 1, wherein the biological sample comprises human, animal, microbial or plant material. 13. The method of claim 1, wherein the magnetic matrix material comprises magnetic beads of silicone, porcelain, ceramic, plastic, glass or polymer. 14. The method of claim 1, wherein the lysis buffer inactivates nucleases, stabilizes macromolecules, and sterilizes the sample at ambient temperatures. 15. The method of claim 1, wherein the one or more specific nucleotide sequences comprises a cancer marker, a sequence indicating the presence of a pathogenic organism or infection, a sequence that is characteristic of a phenotypic condition, a sequence indicating a lineage, a sequence indicating an identifiable characteristic, a sequence indicating a mutation, a sequence indicating a change from a wild-type sequence, or a combination thereof. 16. The method of claim 1, wherein the presence of the specific nucleotide sequence is indicative of the presence of a pathogen. 17. The method of claim 16, wherein the pathogen is a virus, a bacterium, a parasite or a fungus. 18. The method of claim 1, wherein the alkaline buffer comprises water, TE, saline, alcohol or a combination thereof. 19. The method of claim 1, wherein the magnetic field is an electro-magnetic field. 20. A method of extracting nucleic acids from a biological sample containing cells and/or microorganisms comprising: combining the biological sample with a lysis buffer to form a mix;combining the mix with magnetic matrix material to form a solution, wherein neither the buffer or matrix material disrupt the cells and/or microorganisms of the biological sample and the magnetic matrix material binds to nucleic acids of the biological sample other than those present within the cells and/or microorganisms;exposing the solution to a magnetic field and removing liquid to concentrate the magnetic matrix material;adding an alkaline buffer to the concentrated magnetic matrix material to form a mixture, wherein the alkaline buffer causes the release of nucleic acids from the magnetic matrix material;adding carboxy-modified magnetic beads in a binding buffer to the mixture wherein the carboxy-modified magnetic beads bind to a specific nucleic acid sequences;exposing the mixture to a magnetic field and removing liquid to isolate the carboxy-modified magnetic beads bound to the specific nucleic acid sequences;eluting the specific nucleic acid sequences from the carboxy-modified magnetic beads using purified water and/or a Tris-EDTA buffer; andidentifying the specific nucleic acid sequences by PCR, wherein the presence of the specific nucleotide sequence in the biological sample is indicative of the presence of a cancer or a pathogen.
연구과제 타임라인
LOADING...
LOADING...
LOADING...
LOADING...
LOADING...
이 특허에 인용된 특허 (147)
Liav Avraham ; Hansjergen Joyce Anne ; Shimasaki Craig David, 4,7-dialkoxy N-acetylneuraminic acid derivatives and methods for detection of influenza type A and B viruses in clinic.
Bridgham John (Palo Alto CA) Geiser Timothy G. (La Honda CA) Hunkapiller Michael W. (San Carlos CA) Kent Stephen B. H. (Pasadena CA) Marriott Mark P. (Los Altos CA) Ramstad Paul O. (Oakland CA) Nordm, Automated polypeptide synthesis apparatus.
Bridgham John (Palo Alto CA) Geiser Timothy G. (La Honda CA) Hunkapiller Michael W. (San Carlos CA) Kent Stephen B. H. (Pasadena CA) Marriott Mark P. (Los Altos CA) Ramstad Paul O. (Oakland CA) Nordm, Automated polypeptide synthesis apparatus.
Bridgham John (Palo Alto CA) Geiser Timothy G. (La Honda CA) Hunkapiller Michael W. (San Carlos CA) Kent Stephen B. H. (Pasadena CA) Marriott Mark P. (Los Altos CA) Ramstad Paul O. (Oakland CA) Nordm, Automated polypeptide synthesis process.
Gayral, Jean Pierre; Picard, Francois; Boissinot, Maurice; Bastien, Martine, Biological reagents and methods to verify the efficiency of sample preparation and nucleic acid amplification and/or detection.
Monthony James F. (Baltimore MD) Stitt David T. (Parkton MD) Gosnell C. Michael (Fallston MD) Stewart Shannon D. (Stewartstown PA), Biological sample collection and transport device.
Bair, Jr., Robert Jackson; Heath, Ellen M.; Meehan, Heather; Paulsen, Kim Elayne; Wages, Jr., John M., Compositions and methods for using a solid support to purify RNA.
Cerutti Peter A. (Poly/Lausanne CHX) Bressoud Albric (Lausanne CHX), Detection of influenza a virus by polymerase chain reaction (PCR) preceded by reverse transcription of a region of the v.
Whiteley Norman M. (San Carlos CA) Hunkapiller Michael W. (San Carlos CA) Glazer Alexander N. (Orinda CA), Detection of specific sequences in nucleic acids.
Doyle Michael V. (Oakland CA) Newell Arthur D. (Orinda CA) Nunberg Jack H. (Oakland CA) White Thomas J. (Oakland CA), Human IL-2 as a vaccine adjuvant.
Hunter Kenneth W. (4401 Dresden St. Kensington MD 20895) Fischer Gerald W. (10748 Wayridge Dr. Gaithersburg MD 20879), Human monoclonal antibody reactive with polyribosylribitol phosphate.
Heller Michael J. (Encinitas CA), Hybridization of polynucleotides conjugated with chromophores and fluorophores to generate donor-to donor energy transfe.
Sette, Alessandro; Sidney, John; Southwood, Scott; Vitiello, Maria A.; Livingston, Brian D.; Celis, Esteban; Kubo, Ralph T.; Grey, Howard M.; Chesnut, Robert W., Inducing cellular immune responses to hepatitis B virus using peptide and nucleic acid compositions.
Liav Avraham ; Shimasaki Craig D. ; Maher James F. ; Clinkscales C. Worth ; Roark Michael D., Kit for visually detecting the presence of a clinical specimen.
Yagi Kunio (Aichi-ken JPX) Noda Hitoshi (Aichi-ken JPX) Ohishi Nobuko (Gifu JPX) Kurono Masayasu (Mie-ken JPX), Liposome for entrapping gene, liposomal preparation and process for the manufacture of the preparation.
Liav Avraham (Denver CO) Maher James F. (Broken Arrow OK) Shimasaki Craig D. (Tulsa OK) Clinkscales C. Worth (Tulsa OK) Roark Michael D. (Owasso OK), Method for visually detecting the presence of a virus in a clinical specimen.
Reece Phillip A.,AUX ; Wu Wen-Yang,AUX ; Jin Betty,AUX ; Krippner Guy Y.,AUX ; Watson Keith Geoffrey,AUX, Method of detection of influenza virus and compounds for use therein.
Turner Gregory A. (Independence MO) Maher James F. (Broken Arrow OK) Clinkscales C. Worth (Tulsa OK) Roark Michael D. (Owasso OK), Methods for diagnosing human influenza and 4-position modified chromogenic N-acetylneuraminic acid substrated for use th.
Fischer, Gerald W.; Schuman, Richard F.; Wong, Hing; Stinson, Jeffrey R., Opsonic and protective monoclonal and chimeric antibodies specific for lipoteichoic acid of gram positive bacteria.
Fischer, Gerald W.; Schuman, Richard F.; Wong, Hing; Stinson, Jeffrey R., Opsonic and protective monoclonal and chimeric antibodies specific for lipoteichoic acid of gram positive bacteria.
Monthony James F. (Baltimore MD) Stitt David T. (Parkton MD) Gosnell C. Michael (Fallston MD) Stewart Shannon D. (Stewartstown PA), Polyurethane biological sample collection and transport device and its use.
Mullis Kary B. (La Jolla CA) Erlich Henry A. (Oakland CA) Arnheim Norman (Woodland Hills CA) Horn Glenn T. (Emeryville CA) Saiki Randall K. (Richmond CA) Scharf Stephen J. (Berkeley CA), Process for amplifying, detecting, and/or cloning nucleic acid sequences.
Mullis Kary B. (Kensington CA) Erlich Henry A. (Oakland CA) Arnheim Norman (Woodland Hills CA) Horn Glenn T. (Emeryville CA) Saiki Randall K. (Richmond CA) Scharf Stephen J. (Berkeley CA), Process for amplifying, detecting, and/or-cloning nucleic acid sequences.
Boom Willem R. (Amsterdam NLX) Adriaanse Henritte M. A. (Arnhem NLX) Kievits Tim (The Hague NLX) Lens Peter F. (Amsterdam NLX), Process for isolating nucleic acid.
Fell ; Jr. H. Perry (Redmond WA) Folger-Bruce Kim R. (Seattle WA) Yarnold Susan M. (Seattle WA), Production of chimeric antibodies by homologous recombination.
Crawford Jack T. (Atlanta GA) Eisenach Kathleen D. (Little Rock AR) Cave M. Donald (Little Rock AR) Bates Joseph H. (Little Rock AR), Repetitive DNA sequence specific for mycobacterium tuberculosis to be used for the diagnosis of tuberculosis.
Liversidge Gary G. (West Chester PA) Cundy Kenneth C. (Pottstown PA) Bishop John F. (Rochester NY) Czekai David A. (Honeoye Falls NY), Surface modified drug nanoparticles.
Judd Amrit K. (Belmont CA) Bucher Doris J. (New York NY) Popple Steven W. (Brooklyn NY), Synthetic peptides for diagnosis and prevention of influenza virus infection and their use.
Picard,Francois J.; Menard,Christian, Universal method and composition for the rapid lysis of cells for the release of nucleic acids and their detection.
van Scharrenburg, Gustaaf J. M.; Brands, Rudi; de Haan, Lolke; Verweij, Willem Ronald; Wilschut, Jan C.; Agsteribbe, Etienne, Vaccines with an LTB adjuvant.
※ AI-Helper는 부적절한 답변을 할 수 있습니다.