System for releasing and isolating nucleic acids
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12Q-001/68
C12N-015/00
C12N-015/63
C12N-001/20
C07H-021/04
출원번호
US-0558038
(2000-04-26)
우선권정보
DE-0012368 (1995-04-01)
발명자
/ 주소
Bienhaus, Gerhard
Schubert, Ulrich
Kolb, Uwe
Stolz, Burkhard
Pasch, Manfred
출원인 / 주소
Roche Diagnostics GmbH
대리인 / 주소
Arent Fox
인용정보
피인용 횟수 :
71인용 특허 :
9
초록▼
A procedure for the release and isolation of nucleic acids from biological compartments of a sample always uses an instrument that can hold one or more sample processsing vessels, maintain the sample processing vessels at a constant temperature, shake the sample processing vessels and separate magne
A procedure for the release and isolation of nucleic acids from biological compartments of a sample always uses an instrument that can hold one or more sample processsing vessels, maintain the sample processing vessels at a constant temperature, shake the sample processing vessels and separate magnetic particles by means of magnetic force. This system greatly simplifies the isolation of nucleic acids.
대표청구항▼
1. A method of isolating nucleic acids from biological compartments of a fluid sample comprising the steps of:incubating the sample in a sample processing vessel with magnetic particles which magnetic particles are capable of binding with the biological compartments; positioning at least one magnet
1. A method of isolating nucleic acids from biological compartments of a fluid sample comprising the steps of:incubating the sample in a sample processing vessel with magnetic particles which magnetic particles are capable of binding with the biological compartments; positioning at least one magnet towards the sample processing vessel to hold the magnetic particles against an inside wall of the sample processing vessel by magnetic force; removing the remaining fluid, from which the biological compartments have been separated, from the sample processing vessel; introducing a second fluid into the sample processing vessel; resuspending the magnetic particles in the second fluid by eliminating the magnetic force which held the magnetic particles against the inside wall of the sample processing vessel, and shaking the sample processing vessel in the absence of the magnetic force; thereafter lysing the biological compartments to form a lysis mixture; and isolating the nucleic acids from the lysis mixture. 2. The method of claim 1, wherein essentially all of the magnetic particles have a diameter of 2.8 μm to 200 μm.3. The method of claim 2, wherein the average magnetic particle size diameter is about 10 μm to 15 μm.4. The method of claim 1, wherein the isolation step comprises immobilizing the nucleic acids on the magnetic particles.5. The method of claim 1, wherein the nucleic acids to be isolated are transferred to another vessel which is configured to receive a pipette.6. The method of claim 1, wherein the magnetic force is eliminated by separating by a sufficient distance the at least one magnet from the outside wall of the sample processing vessel.7. The method of claim 1, wherein the magnetic force is eliminated by positioning a μ-metal between the vessel and the at least one magnet.8. The method of claim 1, wherein each magnet has a mass of about 0.5 g to about 5 g.9. The method of claim 1, wherein each magnet has a mass of about 1 g to about 4 g.10. The method of claim 1, wherein the processing vessel containing the sample is shaken during at least a portion of the incubation step to facilitate binding.11. The method of claim 1, wherein the magnetic force is eliminated and the sample processing vessel is shaken simultaneously.12. The method of claim 1, wherein the steps of positioning at least one magnet near an outside wall of the sample processing vessel such that it holds the magnetic particles against an inside wall of the sample processing vessel, removing the remaining fluid, from which the biological compartments have been separated, from the sample processing vessel, and resuspending the magnetic particles in a second fluid by eliminating the magnetic force which held the magnetic particles against the inside wall of the sample processing vessel, and shaking the sample processing vessel, are repeated until the biological compartments have reached a desired level of purity.13. The method of claim 1, wherein the fluid sample is a body fluid.14. The method of claim 1, wherein the fluid is blood, saliva or urine.15. The method of claim 1, wherein the nucleic acids are isolated by warming the lysis mixture for a sufficient period of time so as to lyse or partially or completely decompose cell walls of the biological compartments and release the nucleic acids contained in the biological compartments, and cooling the lysis mixture under conditions that make it possible to isolate or hybridize the nucleic acids to be isolated or detected.16. The method of claim 15, wherein the lysis mixture is warmed to a temperature above room temperature.17. The method of claim 16, wherein the lysis mixture is warmed to a temperature of about 70° to about 95 ° C.18. The method of claim 1, wherein the nucleic acids are present in the sample reaction vessel throughout the removing, resuspending and lysing steps.19. The method of claim 1, wherein the removing, resuspending and lysing steps take place within a reaction block.20. The method of claim 19, wherein the reaction vessels remain in the reaction block during the removing, resuspending, and lysing steps.21. The method of claim 1, further comprising the step of detecting the nucleic acids.22. A method of isolating nudeic acids from biological compartments of a fluid sample comprising the steps of:incubating the sample in a sample processing vessel with magnetic particles which magnetic particles are capable of binding with the biological compartments; positioning at least one magnet towards the sample processing vessel to hold the magnetic particles against an inside wall of the sample processing vessel by magnetic force; removing the remaining fluid, from which the biological compartments have been separated, from the sample processing vessel; introducing a second fluid into the sample processing vessel; resuspending the magnetic particles in the second fluid by eliminating the magnetic force which held the magnetic particles against the Inside wall of the sample processing vessel, and shaking the sample processing vessel in the absence of the magnetic force; thereafter lysing the biological compartments to form a lysis mixture; and warming the lysis mixture; and cooling the lysis mixture under conditions that make it possible to isolate or hybridize the nucleic acids to be isolated or detected. 23. The method of claim 22, further comprising shaking the sample processing vessel during the incubating to facilitate the binding of magnetic particles to the biological compartments.24. The method of claim 1, wherein the sample processing vessel has an opening at the bottom from which fluid can exit.
연구과제 타임라인
LOADING...
LOADING...
LOADING...
LOADING...
LOADING...
이 특허에 인용된 특허 (9)
Knobel Rolf,CHX, Analytical device for separating magnetic microparticles from suspensions.
Liberti Paul A. (Churchville PA) Wang Yuzhou (Wayne PA) Tang Weixin (Devon PA) Feeley Brian P. (Easton PA) Gohel Dhanesh I. (Philadelphia PA), Apparatus and methods for magnetic separation featuring external magnetic means.
Fujiwara Koichi (Mito JPX) Mizutani ; deceased Hiromichi (late of Tokyo JPX by Hiroko Mizutani ; legal representative), Device for collecting or preparing specimens using magnetic micro-particles.
Kausch Albert P. (Stonington CT) Narayanswami Sandya (Irvine CA) Manning Jerry E. (San Clemente CA) Hamkalo Barbara A. (Laguna Beach CA), Isolation of biological materials using magnetic particles.
Reeve Michael A. (Henley-on-Thames GB2), Method to isolate macromolecules using magnetically attractable beads which do not specifically bind the macromolecules.
Wilson, Brian D.; Anderson, David L.; Davis, Matthew S.; Erickson, Matthew D.; Johnson, Alan N.; Maurer, Garrick A.; Rosen, Michael J.; Sauerburger, Mark F.; Schmidt, Daniel R.; Wiltsie, Joshua D., Assay cartridge with reaction well.
Ammann, Kelly G.; Burns, Ralph E.; Hansberry, Ernest V.; Horner, Glenn A.; Jakub, Cheryl A.; Kling, John E.; Nieglos, Donald J.; Schneider, Robert E.; Smith, Robert J., Automated analyzer for performing a nucleic acid-based assay.
Ochranek, Brian L.; Arnquist, David C.; Oonuma, Takehiko; Tahara, Hirotoshi; Sato, Naoto; Smith, Bradley P., Automated diagnostic analyzers having rear accessible track systems and related methods.
Ochranek, Brian L.; Arnquist, David C.; Oonuma, Takehiko; Tahara, Hirotoshi; Sato, Naoto, Automated diagnostic analyzers having vertically arranged carousels and related methods.
Ochranek, Brian L.; Arnquist, David C.; Oonuma, Takehiko; Tahara, Hirotoshi; Sato, Naoto, Automated diagnostic analyzers having vertically arranged carousels and related methods.
Ammann, Kelly G.; Burns, Ralph E.; Hansberry, Ernest V.; Horner, Glenn A.; Jakub, Cheryl A.; Kling, John E.; Nieglos, Donald J.; Schneider, Robert E.; Smith, Robert J., Automated process for detecting the presence of a target nucleic acid in a sample.
Ammann, Kelly G.; Burns, Ralph E.; Hansberry, Ernest V.; Horner, Glenn A.; Jakub, Cheryl A.; Kling, John E.; Nieglos, Donald J.; Schneider, Robert E.; Smith, Robert J., Automated process for detecting the presence of a target nucleic acid in a sample.
Ammann, Kelly G.; Burns, Ralph E.; Hansberry, Ernest V.; Horner, Glenn A.; Jakub, Cheryl A.; Kling, John E.; Nieglos, Donald J.; Schneider, Robert E.; Smith, Robert J., Automated process for detecting the presence of a target nucleic acid in a sample.
Ammann,Kelly G.; Burns,Ralph E.; Hansberry,Ernest V.; Horner,Glenn A.; Jakub,Cheryl A.; Kling,John E.; Nieglos,Donald J.; Schneider,Robert E.; Smith,Robert J., Automated process for detecting the presence of a target nucleic acid in a sample.
Lair, Gary D.; Nguyen, Thanh N.; Li, Haitao; Li, Florence; Knight, Byron J.; Heinz, Robert E.; Macioszek, Jerzy A.; Davis, Christopher B.; Scalese, Robert F., Continuous process for performing multiple nucleic acid amplification assays.
Hillebrand, Timo; Arndt, Matthias; Wellnitz, Uwe; Berka, Klaus; Hillebrand, Volker, Device and procedure for automated isolation and purification of nucleic acids from complex starting materials of the users choice.
Ammann, Kelly G.; Schneider, Robert E.; Smith, Robert J., Method for agitating the contents of a reaction receptacle within a temperature-controlled environment.
Macioszek, Jerzy A.; Davis, Christopher B.; Lair, Gary D.; Nguyen, Thanh N.; Li, Haitao; Li, Florence F.; Knight, Byron J.; Scalese, Robert F.; Heinz, Robert E., Method for continuous mode processing of the contents of multiple reaction receptacles in a real-time amplification assay.
Ammann, Kelly G.; Burns, Ralph E.; Hansberry, Ernest V.; Horner, Glenn A.; Jakub, Cheryl A.; Kling, John E.; Nieglos, Donald J.; Schneider, Robert E.; Smith, Robert J., Method for detecting the presence of a nucleic acid in a sample.
Ammann, Kelly G.; Burns, Ralph E.; Hansberry, Ernest V.; Horner, Glenn A.; Jakub, Cheryl A.; Kling, John E.; Nieglos, Donald J.; Schneider, Robert E.; Smith, Robert J., Method for detecting the presence of a nucleic acid in a sample.
Ammann, Kelly G.; Burns, Ralph E.; Hansberry, Ernest V.; Horner, Glenn A.; Jakub, Cheryl A.; Kling, John E.; Nieglos, Donald J.; Schneider, Robert E.; Smith, Robert J., Method for detecting the presence of a nucleic acid in a sample.
Ammann, Kelly G.; Burns, Ralph E.; Hansberry, Ernest V.; Horner, Glenn A.; Jakub, Cheryl A.; Kling, John E.; Nieglos, Donald J.; Schneider, Robert E.; Smith, Robert J., Method for detecting the presence of a nucleic acid in a sample.
Ammann, Kelly G.; Schneider, Robert E.; Smith, Robert J., Method for introducing a fluid into a reaction receptacle contained within a temperature-controlled environment.
Ammann, Kelly G.; Burns, Ralph E.; Hansberry, Ernest V.; Horner, Glenn A.; Jakub, Cheryl A.; Kling, John E.; Nieglos, Donald J.; Schneider, Robert E.; Smith, Robert J., Method for performing an assay with a nucleic acid present in a specimen.
Macioszek, Jerzy A.; Davis, Christopher B.; Lair, Gary D.; Nguyen, Thanh N.; Li, Haitao; Li, Florence F.; Knight, Byron J.; Scalese, Robert F.; Heinz, Robert E., Method for performing multi-formatted assays.
Ammann, Kelly G.; Burns, Ralph E.; Hansberry, Ernest V.; Horner, Glenn A.; Jakub, Cheryl A.; Kling, John E.; Nieglos, Donald J.; Schneider, Robert E.; Smith, Robert J., Method for simultaneously performing multiple amplification reactions.
Knight, Byron J.; Opalsky, David; Schroeter, Brian, Method, system and apparatus for incorporating capacitive proximity sensing in an automated fluid transfer procedure.
Wilson, Brian D.; Davis, Matthew S.; Erickson, Matthew D.; Johnson, Alan N.; Maurer, Garrick A.; Schmidt, Daniel R.; Wiltsie, Joshua D., Reaction vessel.
Lair, Gary D.; Nguyen, Thanh N.; Li, Haitao; Li, Florence F.; Knight, Byron J.; Heinz, Robert E.; Macioszek, Jerzy A.; Davis, Christopher B.; Scalese, Robert F., Signal measuring system having a movable signal measuring device.
Wilson, Brian D.; Anderson, David L.; Erickson, Matthew D.; Johnson, Alan N.; Rosen, Michael J.; Schmidt, Daniel R.; Wiltsie, Joshua D., System and method including analytical units.
Wilson, Brian D.; Anderson, David L.; Davis, Matthew S.; Erickson, Matthew D.; Johnson, Alan N.; Maurer, Garrick A.; Rosen, Michael J.; Sauerburger, Mark F.; Schmidt, Daniel R.; Wiltsie, Joshua D., System and method including multiple processing lanes executing processing protocols.
Wilson, Brian D.; Alaruri, Sami D.; Davis, Matthew S.; Erickson, Matthew D.; Johnson, Alan N.; Maurer, Garrick A.; Sauerburger, Mark F.; Schmidt, Daniel R.; Wiltsie, Joshua D.; Stachelek, Thomas M.; Yang, David L., System and method including thermal cycler modules.
Lair, Gary D.; Nguyen, Thanh N.; Li, Haitao; Li, Florence F.; Knight, Byron J.; Heinz, Robert E.; Macioszek, Jerzy A.; Davis, Christopher B.; Scalese, Robert F., System for performing multi-formatted assays.
Heinz, Robert E.; Newell, Dennis; Opalsky, David; Rhubottom, Jason, Systems and methods for distinguishing optical signals of different modulation frequencies in an optical signal detector.
Heinz, Robert E.; Newell, Dennis; Opalsky, David; Rhubottom, Jason, Systems and methods for distinguishing optical signals of different modulation frequencies in an optical signal detector.
※ AI-Helper는 부적절한 답변을 할 수 있습니다.