WO2003016558A1 - Continuous flow thermal device - Google Patents
Continuous flow thermal device Download PDFInfo
- Publication number
- WO2003016558A1 WO2003016558A1 PCT/AU2002/001112 AU0201112W WO03016558A1 WO 2003016558 A1 WO2003016558 A1 WO 2003016558A1 AU 0201112 W AU0201112 W AU 0201112W WO 03016558 A1 WO03016558 A1 WO 03016558A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- fluid
- conduit
- temperature
- heat exchanger
- reaction
- Prior art date
Links
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
- B01L7/525—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples with physical movement of samples between temperature zones
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/08—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/02—Adapting objects or devices to another
- B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/04—Closures and closing means
- B01L2300/041—Connecting closures to device or container
- B01L2300/044—Connecting closures to device or container pierceable, e.g. films, membranes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1838—Means for temperature control using fluid heat transfer medium
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N2021/7769—Measurement method of reaction-produced change in sensor
- G01N2021/7786—Fluorescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
Definitions
- the present invention relates to a device which is able to control and vary in a cyclical manner, or in a progressive manner, the temperature of a fluid in a tube or is able to maintain fluid in a tube at a constant temperature; and is particularly applicable to monitoring the course of a reaction by real time scanning. It is applicable to a device for amplifying nucleic acids when amplification takes place by use of reactions such as the polymerase chain reaction, the ligase chain reaction and the like, when the device is used to vary temperature; and is also applicable to isothermal reactions when the device maintains the temperature at a constant level.
- the present invention when it relates to a temperature cycling device, takes reactants through different temperatures in a cyclical fashion where it is necessary that the temperature change be achieved accurately and rapidly.
- the present invention provides a system where the reaction, whether carried out in a cyclical manner, a progressive manner, or isothermally, can be monitored by real time monitoring. For example, in the case of cyclic reactions, it is possible to monitor the course of a reaction at each cycle.
- PCR polymerase chain reaction
- thermostable nucleic acid polymerases changed the situation dramatically. These enzymes are stable at about 95°C and therefore it was not necessary to replace these enzymes after heating to separate the two strands of DNA. Thus, devices which took advantage of a thermostable nucleic acid polymerase were developed.
- a thermal cycling device including: a heat exchanger body having a longitudinal axis and longitudinally divided to provide at least two segments which are able to be heated to different temperatures so that said body has peripheral surfaces of different temperatures; a conduit extending about said body so as to be in thermal contact with said peripheral surfaces; a first delivery device to deliver a first fluid to said conduit to cause said fluid to pass therealong and therefore change in temperature as the fluid passes said segments; and a second delivery device to deliver a second fluid to said conduit so as to flow with said first liquid and therefore also change in temperature.
- the body is longitudinally divided so that said segments and surfaces are longitudinally extending, and said conduit extends angularly about said body relative to said axis.
- PCR polymerase chain reaction
- LCR ligase chain reaction
- the first fluid comprises a sample to be analysed and reagents used in the analysis and the second fluid comprises a transport fluid.
- the sample to be analysed is a nucleic acid and may be DNA or RNA.
- the reagents used in the analysis are well known in the art, including for example nucleic acid polymerase, for example Taq polymerase.
- a thermal cycling device including: a heat exchanger body having a longitudinal axis and transversely divided to provide at least two segments which are able to be heated to different temperatures so that said body has peripheral surfaces of different temperatures; a conduit extending about said body so as to be in thermal contact with said peripheral surfaces; a first delivery device to deliver a first fluid to said conduit to cause said fluid to pass therealong and therefore change in temperature as the fluid passes said segments; and a second delivery device to deliver a second fluid to said conduit so as to flow with said first liquid and therefore also change in temperature.
- the body is transversely divided relative to said axis so that said segments and surfaces extend angularly about said axis, and said conduit extends longitudinally of said body relative to said axis.
- This aspect is applicable where the temperature is changed in a progressive manner from one temperature to another temperature, for example, to DNA melt detection studies.
- the first fluid comprises a sample to be analysed and the second fluid comprises a transport fluid.
- the sample to be analysed is a nucleic acid and may be DNA or RNA.
- the reagents used in the analysis are well known in the art.
- an isothermal device including: a heat exchanger body having a longitudinal axis and a peripheral surface and means for maintaining the temperature of said peripheral surface at a constant selectable temperature; a conduit extending about said body so as to be in thermal contact with said peripheral surface; a first delivery device to deliver a first fluid to said conduit to cause said fluid to pass therealong and therefore change in temperature as the fluid passes said peripheral surface; and a second delivery device to deliver a second fluid to said conduit so as to flow with said first liquid and therefore also change in temperature.
- the heat exchanger may be a solid body which can maintain a fixed, selectable temperature.
- the first fluid comprises an antibody and antigen to said antibody and the second fluid comprises a transport fluid.
- conduit which extends about the heat exchanger body in the three aspects above is disposed in a series of consecutive windings and that the heat exchanger body is solid.
- the device of each embodiment also comprises a scanning detector which is able to monitor the progress of a chemical reaction occurring in the conduit.
- the scanning detector is further described below. With the addition of this scanning detector, it is possible, in each of the above- described embodiments to monitor the course of a reaction at any point on the outside of the thermal cycling device or isothermal device.
- the heat exchanger body is in the form of a cylinder or equivalent shape, it may be divided into "pie" shaped segments each being at a different temperature (typically about 94°C, 55°C, 72°C) so that as the samples migrate around one turn of the spiral, the temperature in the fluid is cycled to those temperatures corresponding to the "pie" segment of the body.
- the heat exchanger body may have a temperature gradient applied across it from one end to the opposite end.
- the temperature gradient would be set from one flat surface to the opposite flat surface to the top flat surface so that as the samples migrate around the spiral the temperature is gradually increased or decreased.
- the heat exchanger body may also be maintained at a uniform temperature, where the temperature may suitably be maintained at any temperature from 0-100°C.
- Each of the different temperatures may be achieved by standard electrical heating and optionally air cooling to adjust the temperature; by circulating fluid at a particular temperature; or by the use of Peltier devices. It is particularly preferred that Peltier devices be used since the use of such devices allows a more accurate temperature selection.
- An important aspect of temperature control in the first aspect is based on the fact that fluid in the conduit passes from a segment at 95°C to a segment at 55°C. The effect of this is that in the 55°C segment, fluid would tend to heat the surface of the 55°C segment. It is for this reason that air cooling may be necessary depending on the speed of fluid in the conduit and hence transfer of heat from the fluid back to the segment at the lower temperature of 55°C.
- the conduit is preferably disposed in the form of a series of coils arranged in a spiral fashion around the cylinder.
- the device of the first embodiment is particularly applicable and each turn of the spiral represents one cycle in the PCR and it is therefore possible to choose the number of cycles by equating this to the number of turns of conduit.
- the number of turns is 35-60.
- the device comprises 60 turns which in practice allows a choice of scanning as many turns as required.
- the back pressure which is applied should be between about 300 and about 700kPa and may be achieved by methods known in the art. For example, it may be achieved by use of a spring and ball device at the fluid outlet where adjusting the tension of the spring sets the pressure.
- the conduit is suitably tubing which has the following properties: (1) It is able to conduct heat from the heat exchanger body to fluid in the conduit efficiently so that a change in temperature in a particular portion of the heat exchanger body accurately causes a change in temperature in the conduit adjacent that portion of the heat exchanger body which in turn causes a change in temperature in the fluid inside the corresponding interior portion of the conduit. (2) The interior surface of the conduit is hydrophobic so that the reaction mixture has no affinity for the interior surface of the tubing.
- the conduit is PTFE tubing or similar.
- the fluid present in the conduit is any fluid where the temperature needs to be controlled precisely and accurately and either cyclically, progressively or isothermally.
- the fluid comprises a reaction mixture containing reagents to amplify one or more nucleic acids and a sample containing one or more nucleic acids.
- the nucleic acids to be amplified are either RNA or DNA and the enzymes used in the reaction mixture are any enzymes well known in this art.
- DNA polymerases may suitably be used to amplify DNA and preferably thermostable DNA polymerases are particularly applicable in the reactions carried out in the device of this invention.
- the fluid may be divided into two streams as set out in the first embodiment of the invention described above. One such fluid contains samples to be analysed while the other fluid contains reagents used in the analysis.
- the conduit is also filled with a carrier fluid which is any synthetic based oil that is completely free of
- RNA or DNA typically, silicon oil is suitable for use in this device.
- silicon oil is suitable for use in this device.
- the reaction mixture is run through the conduit under a back pressure to stop the degassing of samples during the heating process.
- each reaction mixture is sequentially injected into the carrier fluid at regular spacings.
- spacing between samples is suitably achieved by setting a distance which is approximately 5 times the internal diameter of the tube between reactants.
- the means for causing fluid to be transported through the conduit is for example any suitable pump means. This may comprise a peristaltic pump, for example. It is important that such a means for pumping fluid through the conduit is a precision pump for the reasons described below.
- the heat exchanger body is any body which is so shaped that it can conveniently accommodate a conduit disposed thereon. It is preferably substantially cylindrically shaped. However, any generally regular, three dimensional shaped body which can accommodate conduit to effectively transfer heat to a fluid in the conduit would be acceptable, provided that its shape permits disposition of conduit on the surface of the heat exchanger in a regular, repeating fashion, each repeat representing one temperature cycle in the case of the device being used in a cyclic reaction.
- the heat exchanger body has a groove machined into the surface accommodating the conduit and is dimensioned such that it can accommodate the conduit in a close fitting manner.
- the reason for this groove is to increase the efficiency of heat transfer from the heat exchanger body to the conduit and hence to the fluid therein.
- this device for example the PCR and LCR, it is vitally important that ideally no cross contamination between samples occurs. Even if some contamination between samples does occur it is important that this be kept to an absolute minimum. In order to achieve no or minimal contamination, there is provided an injector means.
- an injector means for use in a thermal cycling device comprising: a septum; a needle which is able to pierce the septum; a reservoir in fluid contact with the septum, having an inlet and an outlet, and able to accommodate a purge fluid which is able to enter the reservoir via the inlet and leave the reservoir via the outlet; wherein when in use, the needle passes through fluid in said reservoir and then through the septum.
- the injector means comprises a septum and is suitably a rubber septum of the type used in gas chromatography.
- Another component of the injector is a needle which pierces the septum. Cross contamination is prevented by filling a reservoir above the septum with purge fluid, urging the needle through the reservoir of fluid then through the septum while continually purging the area with purge fluid, withdrawing the needle and finally purging the withdrawn needle prior to its subsequent penetration of the septum with the next sample.
- Purge fluid may be silicon oil or PCR grade water or any synthetic oil which is devoid of RNA or DNA.
- the septum is made of material which closes about the hole made by a previous penetration. It is preferably made of rubber that is, silicon rubber or natural rubber and is more preferably the same material used in gas chromatography. It is also important, that when back pressure is applied, the septum is of a material which can withstand such back pressure. Any carry over from the last sample will potentially result in a false positive in the following sample. To overcome this, a negative control is run between each of the samples. Typically, this could be 1 in every 10 samples depending on the characteristics of a particular assay.
- the entire system could then the purged, both the loading port and the entire spiral tubing, and the last 10 samples (or whatever is considered necessary by the operator) could be re-run and the job continued without intervention.
- the system could also be flushed with NaOH to degrade any DNA in the system.
- the first fluid is delivered by a first fluid device, that is the injector means described above.
- the second fluid, transport fluid is delivered by a second fluid device.
- the transport fluid may be any fluid which is inert and does not interfere with the analysis and furthermore is devoid of DNA and RNA.
- the transport fluid may be an inert oil and is typically silicon oil. It is preferred that the transport fluid is delivered under pressure, typically between about 700 and 7000 kPa.
- back pressure is suitably between about 300 and about 700kPa and is achieved by methods known in the art. Specifically, it may be achieved by use of a spring and ball device at the outlet wherein adjusting the tension of the spring sets the pressure. The effect of this is to prevent vaporisation of the fluid stream and reaction mixtures as they pass through the zones of high temperature. As also described in our earlier patent, in order to prevent mixing of reaction mixtures, it is preferred that the flow rate and cross-sectional diameter of the fluid stream are such that turbulent flow is avoided and laminar flow is achieved.
- tubing of small diameter for example about 3mm (or 1/8 inch) is particularly preferred since it is both beneficial to rapid heat transfer and minimises turbulent flow.
- the tube diameter could also be decreased to reduce the reaction volume from 20 ⁇ L to l-2 ⁇ L. In this instance the diameter would be about 0.5 mm outside diameter and about 0.2 mm (inside diameter).
- the samples travel along the conduit at a constant velocity (regulated by the precision pump), and so take a known amount of time to complete one revolution. Typically it takes about 1 minute to make a full revolution, and based on a 35 coil system a sample would take 35 minutes to completely flow through the device.
- a typical velocity of fluid is 5mm per second, with a diameter of 100mm the circumference is approx 300mm and so a sample takes approx 1 minute for a revolution.
- a typical sample volume is 20-50 ⁇ L (which, as mentioned above, may be reduced to 1-2 ⁇ L) and the length of a sample in the conduit is 5-lOmm. Given that at least 10 readings of the same sample should be taken by the scanner (to be described presently) to obtain an average fluorescent energy, the scan head must scan all coils (typically, 35 coils) in 0.5 seconds. The maximum flow rate will therefore be limited by the scan rate of the scanning head.
- a scanning detector in the case of the heat exchanger body being cylindrical, is fitted to the outside of the cylinder and runs from the first coil to the last coil. As the scanning head of the detector moves past each of the coils it detects the fluorescence energy in the conduit. If a sample is present it detects the sample energy, and if only the carrier fluid is present it detects only the background energy of the carrier fluid.
- the scanning head may be manufactured as an integrated part of the system and multiple heads may be attached to detect up to 4 channels.
- the heat exchanger body itself may be rotated relative to the scanning head. This would allow readings to be taken at different temperature zones by rotating the conduits past the reaction head.
- the scanning head suitably consists of a mirror set to direct a laser beam onto the coils, and an optic fibre to collect the fluorescent energy which pipes the collected light to a photomultiplier tube that converts light to a voltage captured by a computer system. On every tenth loading a tracking dye is injected to give a full scale signal so samples order can be maintained in the software.
- a commercially available scanning head which would be acceptable would be a design similar to the head used in the GS-2000 Gel-Scan.
- a light source laser or light emitting diodes
- the tubing is substantially transparent to fluorescent light emitted from the samples.
- fluorescent dyes are for example, FAM, JOE, and ROX, which are all trademarks of ABI or Cy5.
- any reaction which may be coupled with a reagent where colour change indicates the progress of a reaction would be suitable.
- chromogenic substrates where suitable, for example chromogenic substrates used in coagulation tests would be suitable.
- the device described typically enables a sample to be loaded every 5 seconds into the fluid carrier stream, this equates to automated loading of 18,000 samples in a 24 hour period.
- the device of this invention therefore has 3 times the throughput, none of the associated robotics for transporting and sealing of plates, and therefore none of the problems associated with such systems.
- a separate heating device may be added which is placed before the heat exchanger body described above.
- this may also comprise a cylinder on which the conduit is disposed so that the fluid may be optionally heated prior to its transport to the heat exchanger of the present invention.
- Figure 1 is a schematic representation of a cylindrically shaped heat exchanger in accordance with the first aspect of this invention used in the PCR together with a schematic representation of the scanner;
- Figure 2A is a schematic representation of the "pie" shaped segments of the heat exchanger body of Figure 1;
- Figure 2B is a schematic representation of an alternative scanning arrangement of
- Figure 3 is a cross section of the injector means
- Figure 4 is a schematic representation of a cylindrically shaped heat exchanger in accordance with the second aspect of this invention
- Figure 5 shows the results of a DNA melt experiment using the device schematically represented in Figure 4;
- Figure 6 is a schematic representation of a cylindrically shaped isothermal device in accordance with the third aspect of this invention.
- Conduit 1 carries the reaction mixture which is represented as being delivered by the injector means by arrow 35 in Figure 3 to the first coil 2 of the heat exchanger body 3 which in Figure 1 is cylindrically shaped. Heat is transferred from the heat exchanger body 3 to conduit 2 containing the fluid whose temperature then achieves that of the portion of the heat exchanger body adjacent it. The segments of different temperatures are more clearly shown in Figure 2 A and Figure 2B.
- the number of turns of conduit disposed on the heat exchanger body represents the number of cycles of nucleic acid amplification. On completion of the desired number of cycles of the amplification reaction, the reaction fluid exits the device by conduit 4.
- the device may be used preparatively, that is, the fluid may be collected and the amplified nucleic acid isolated; or analytically, that is, the fluid may be analysed.
- a scanning detector 5 is fitted to the outside of the heat exchanger. The direction of the arrow 6 indicates that the scanning head 7 travels past the coils and detects sample energy emanating from those coils.
- reaction mixture represented as being delivered by the injector means by arrow 35 in Figure 3, enters by conduit 10 and firstly encounters a segment 11 held at approximately 95°C. It is then transported to a zone 12 which is held at 55°C at which temperature the amplification reaction takes place. The fluid is then transported to the segment 13 held at approximately 72°C. As can be seen from Figure. 2A the fluid passes the head 14 of the scanner detector 15 as fluid leaves segment 11, held at approximately 95°C.
- FIG. 2B An alternative scanning arrangement is shown in Figure 2B where the heat exchanger body rotates about its longitudinal axis through 180° approximately. This is indicated by double headed arrow 16.
- An advantage of this arrangement is that the same scanning head could monitor the reaction in different temperature zones.
- the scanning head itself may rotate about the longitudinal axis of the heat exchanger body through 180°, also indicated by the double headed arrow 16. This would also allow the reaction to be monitored in different temperature zones.
- the injector means is depicted in Figure. 3. This consists of a needle 20 into which is loaded the reagent mixture comprising reagent 21 and nucleic acid sample 22. This reaction mixture is separated from the preceding mixture and following mixture by carrier fluid 23.
- the reaction mixture When loading the sample into the conduit, the reaction mixture is pumped down the needle by means of a pump (not shown) in the direction of arrow 24 through tip 25.
- a pump Prior to piercing septum 26, the needle is washed by purge fluid which is situated in reservoir 27.
- the purge fluid is introduced into the reservoir 27 by an inlet 28 and is maintained at a height 29 due to outflow of purge fluid through outlet 30 when the needle 20 pierces septum 26, the pump (not shown) expels the contents of needle 20 into the conduit by stopping at position 31 whereupon the contents are expelled into a constantly flowing stream of transport oil which enters conduit 33 at position 32.
- the reaction mixture and transport oil are then transported under the pressure of the oil inlet pump schematically represented at 34.
- the mixture is transported to the heat exchanger body in the direction of arrow 35 and undergoes cycles of amplification on the body represented in Figures 1 and 2.
- the number of cycles is determined by the number of windings of conduit chosen by the operator.
- FIG. 4 A further embodiment of this invention is depicted in Figure. 4.
- the reaction mixture represented as being delivered by the injector means by arrow 35 in Figure 3 enters the device by conduit 44.
- the heat exchanger body includes a solid aluminium rod 40 on which is wound a series of coils of conduit 41.
- the method will be further exemplified in Example 2.
- the temperature at which the reaction mixture in the conduit arrives at the heat exchanger body at end 45 is 50°C and the final temperature at end 46 is 95°C.
- a back pressure is applied at position 43.
- the scanning device 42 includes a laser source 47 which transmits a laser beam 48 to a beam splitter 49.
- the resulting fluorescence caused by the reaction in the conduit is detected by detector 42 and analysed by methods known in the art.
- the scanner is able to move along the outside of the heat exchanger and can monitor the course of the reaction at any point.
- Figure 5 shows a typical DNA melt detection analysis.
- the x-axis represents temperature and the y-axis represents dF/dT which is the change in fluorescence compared to change in temperature.
- the vertical line 50 in the graph represents the left hand side of the device 40 in Figure 4 while the vertical line 51 represents the right hand side of the device 40 in Figure 4.
- Horizontal line 52 in the graph represents the threshold level determined by computer representing signal above the noise level.
- the peak at 69°C (ie reference numeral 53) represents wild type DNA; the peaks at 62°C and 69°C (ie reference numerals 54 and 53 respectively) represent heterozygous DNA; and the peak at 62°C (ie reference numeral 54) represents a mutation in the DNA.
- FIG. 6 illustrates the isothermal device of the third aspect of this invention.
- Conduit 61 carries the reaction mixture which is represented as being delivered by the injector means by arrow 35 in Figure 3 to the first coil 62 of the heat exchanger body 63 which in Figure 6 is cylindrically shaped.
- the fluid which enters conduit 61 consists of a mixture of two fluids namely an antigen and an antibody to the antigen which have been mixed prior to entering conduit 61. Heat is transferred from the heat exchanger body 63 to conduit 62 containing the fluid whose temperature then achieves that of the heat exchanger body.
- reaction fluid exits the device by conduit 64.
- a scanning detector 65 is fitted to the outside of the heat exchanger. The direction of the arrow 66 indicates that the scanning head 67 travels past the coils and is able to monitor the reaction taking place therein.
- the injector means it is not as important as in the first aspect of the invention to prevent cross contamination and therefore, it is not vital that the injector means be used.
- reactions between antigens and antibodies are normally carried out at approximately 45 °C, degassing will not occur and therefore it is not essential to apply a back pressure to the fluid leaving the device.
- the needle is purged with silicon carrier oil.
- (C) Oil Inlet from Pump A high pressure (typically about 700- about 7000kPa, that is about 100- about 1000 psi) constant flow pump is used to regulate the main carrier oil flow through the injector port that is coupled to the real-time flow through heating system.
- a high pressure typically about 700- about 7000kPa, that is about 100- about 1000 psi
- constant flow pump is used to regulate the main carrier oil flow through the injector port that is coupled to the real-time flow through heating system.
- oligo probe typically a 20 mer
- these samples will have a higher melting point.
- the probe hybridizes to a mutated DNA sample, there is a base mismatch and the probe is not 100% homologous and so the thermal energy required to denature the probe from the DNA target is less which produces a lower melting point.
- FIG. 4 schematically illustrates use of the heat exchanger in this method. That is, the heat exchanger body is heated at one end at approximately 50°C and at the other end at approximately 95 °C. The temperature gradually changes over the length of the cylinder. In this instance, 45 turns give approximately 1°C difference. That is, each turn is 1°C hotter than the turn before it.
- Figure 5 is a DNA melt experiment using the apparatus of Figure 4 where the temperature is adjusted in accordance with the second aspect of this invention. The higher temperature peak in Figure. 5 is wild type DNA and the lower temperature peak is mutated DNA Industrial Applicability
Abstract
Description
Claims
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AU2002322178A AU2002322178B2 (en) | 2001-08-16 | 2002-08-16 | Continuous flow thermal device |
US11/149,217 US7709250B2 (en) | 2001-08-16 | 2005-06-10 | Continuous flow thermal device |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AUPR7071A AUPR707101A0 (en) | 2001-08-16 | 2001-08-16 | Continuous flow thermal device |
AUPR7071 | 2001-08-16 |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/149,217 Continuation US7709250B2 (en) | 2001-08-16 | 2005-06-10 | Continuous flow thermal device |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2003016558A1 true WO2003016558A1 (en) | 2003-02-27 |
Family
ID=3831026
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/AU2002/001112 WO2003016558A1 (en) | 2001-08-16 | 2002-08-16 | Continuous flow thermal device |
Country Status (3)
Country | Link |
---|---|
US (1) | US7709250B2 (en) |
AU (2) | AUPR707101A0 (en) |
WO (1) | WO2003016558A1 (en) |
Cited By (42)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005023427A1 (en) * | 2003-09-05 | 2005-03-17 | Stokes Bio Limited | A microfluidic analysis system |
EP1784505A1 (en) * | 2004-09-02 | 2007-05-16 | Bioneer Corporation | Miniaturized apparatus for real-time monitoring |
WO2007091230A1 (en) * | 2006-02-07 | 2007-08-16 | Stokes Bio Limited | A microfluidic analysis system |
FR2907227A1 (en) * | 2006-10-13 | 2008-04-18 | Rhodia Recherches & Tech | METHOD AND FACILITY FOR DETERMINING AT LEAST ONE PARAMETER OF A PHYSICAL AND / OR CHEMICAL TRANSFORMATION AND CORRESPONDING SCREENING METHOD |
EP1982195A1 (en) * | 2006-02-02 | 2008-10-22 | Corbett Life Science Pty Ltd | Thermocycler and sample port |
WO2009157695A3 (en) * | 2008-06-23 | 2010-03-25 | Bioneer Corporation | Thermal cycling reaction block and continuous real-time monitoring apparatus using the same |
US20100120635A1 (en) * | 2003-09-05 | 2010-05-13 | Stokes Bio Limited | Sample dispensing |
USRE41780E1 (en) | 2003-03-14 | 2010-09-28 | Lawrence Livermore National Security, Llc | Chemical amplification based on fluid partitioning in an immiscible liquid |
JP2011513743A (en) * | 2008-03-03 | 2011-04-28 | ロディア オペレーションズ | Method and apparatus for determining at least one parameter of physical and / or chemical transition |
EP2553085A2 (en) * | 2010-03-26 | 2013-02-06 | Stokes Bio Limited | Devices, systems, and methods for amplifying nucleic acids |
US20130040381A1 (en) * | 2004-01-28 | 2013-02-14 | Marshall University Research Corporation | Apparatus and method for a continuous rapid thermal cycle system |
US8550503B2 (en) | 2006-09-28 | 2013-10-08 | Stokes Bio Ltd. | Microfluidic connector |
US8633015B2 (en) | 2008-09-23 | 2014-01-21 | Bio-Rad Laboratories, Inc. | Flow-based thermocycling system with thermoelectric cooler |
US8697011B2 (en) | 2009-05-19 | 2014-04-15 | Stokes Bio Limited | Sampling device with immiscible fluid supply tube in counter-flow arrangement |
US8741660B2 (en) | 2009-05-19 | 2014-06-03 | Stokes Bio Limited | Sampling device |
US8951939B2 (en) | 2011-07-12 | 2015-02-10 | Bio-Rad Laboratories, Inc. | Digital assays with multiplexed detection of two or more targets in the same optical channel |
US9089844B2 (en) | 2010-11-01 | 2015-07-28 | Bio-Rad Laboratories, Inc. | System for forming emulsions |
US9126160B2 (en) | 2008-09-23 | 2015-09-08 | Bio-Rad Laboratories, Inc. | System for forming an array of emulsions |
US9132394B2 (en) | 2008-09-23 | 2015-09-15 | Bio-Rad Laboratories, Inc. | System for detection of spaced droplets |
US9194861B2 (en) | 2009-09-02 | 2015-11-24 | Bio-Rad Laboratories, Inc. | Method of mixing fluids by coalescence of multiple emulsions |
US9222128B2 (en) | 2011-03-18 | 2015-12-29 | Bio-Rad Laboratories, Inc. | Multiplexed digital assays with combinatorial use of signals |
US9347059B2 (en) | 2011-04-25 | 2016-05-24 | Bio-Rad Laboratories, Inc. | Methods and compositions for nucleic acid analysis |
US9393560B2 (en) | 2010-03-25 | 2016-07-19 | Bio-Rad Laboratories, Inc. | Droplet transport system for detection |
US9399215B2 (en) | 2012-04-13 | 2016-07-26 | Bio-Rad Laboratories, Inc. | Sample holder with a well having a wicking promoter |
US9417190B2 (en) | 2008-09-23 | 2016-08-16 | Bio-Rad Laboratories, Inc. | Calibrations and controls for droplet-based assays |
US9492797B2 (en) | 2008-09-23 | 2016-11-15 | Bio-Rad Laboratories, Inc. | System for detection of spaced droplets |
US9500664B2 (en) | 2010-03-25 | 2016-11-22 | Bio-Rad Laboratories, Inc. | Droplet generation for droplet-based assays |
US9597644B2 (en) | 2003-09-05 | 2017-03-21 | Stokes Bio Limited | Methods for culturing and analyzing cells |
US9598725B2 (en) | 2010-03-02 | 2017-03-21 | Bio-Rad Laboratories, Inc. | Emulsion chemistry for encapsulated droplets |
US9764322B2 (en) | 2008-09-23 | 2017-09-19 | Bio-Rad Laboratories, Inc. | System for generating droplets with pressure monitoring |
US9904734B2 (en) | 2013-10-07 | 2018-02-27 | Apdn (B.V.I.) Inc. | Multimode image and spectral reader |
US9963740B2 (en) | 2013-03-07 | 2018-05-08 | APDN (B.V.I.), Inc. | Method and device for marking articles |
US10047282B2 (en) | 2014-03-18 | 2018-08-14 | Apdn (B.V.I.) Inc. | Encrypted optical markers for security applications |
WO2019193104A1 (en) | 2018-04-04 | 2019-10-10 | Altana Ag | Effect pigments based on colored hectorites and coated colored hectorites and manufacture thereof |
US10512910B2 (en) | 2008-09-23 | 2019-12-24 | Bio-Rad Laboratories, Inc. | Droplet-based analysis method |
US10519605B2 (en) | 2016-04-11 | 2019-12-31 | APDN (B.V.I.), Inc. | Method of marking cellulosic products |
US10730051B2 (en) | 2006-02-07 | 2020-08-04 | Stokes Bio Ltd. | Liquid bridge and system |
US10741034B2 (en) | 2006-05-19 | 2020-08-11 | Apdn (B.V.I.) Inc. | Security system and method of marking an inventory item and/or person in the vicinity |
US10745825B2 (en) | 2014-03-18 | 2020-08-18 | Apdn (B.V.I.) Inc. | Encrypted optical markers for security applications |
US10920274B2 (en) | 2017-02-21 | 2021-02-16 | Apdn (B.V.I.) Inc. | Nucleic acid coated submicron particles for authentication |
US10995371B2 (en) | 2016-10-13 | 2021-05-04 | Apdn (B.V.I.) Inc. | Composition and method of DNA marking elastomeric material |
US11130128B2 (en) | 2008-09-23 | 2021-09-28 | Bio-Rad Laboratories, Inc. | Detection method for a target nucleic acid |
Families Citing this family (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2660482B1 (en) * | 2005-08-22 | 2019-08-07 | Life Technologies Corporation | Vorrichtung, System und Verfahren unter Verwendung von nichtmischbaren Flüssigkeiten mit unterschiedlichen Volumen |
US7854902B2 (en) * | 2006-08-23 | 2010-12-21 | Nanotek, Llc | Modular and reconfigurable multi-stage high temperature microreactor cartridge apparatus and system for using same |
US8709762B2 (en) | 2010-03-02 | 2014-04-29 | Bio-Rad Laboratories, Inc. | System for hot-start amplification via a multiple emulsion |
EP2550351A4 (en) | 2010-03-25 | 2014-07-09 | Quantalife Inc | Detection system for droplet-based assays |
US20120034688A1 (en) * | 2010-08-04 | 2012-02-09 | Griffin Stephen E | True nucleic acid amplification |
WO2013019751A1 (en) | 2011-07-29 | 2013-02-07 | Bio-Rad Laboratories, Inc., | Library characterization by digital assay |
US9222115B2 (en) | 2011-12-30 | 2015-12-29 | Abbott Molecular, Inc. | Channels with cross-sectional thermal gradients |
US20140255270A1 (en) * | 2013-02-28 | 2014-09-11 | California Institute Of Technology | Removing sacrificial layer to form liquid containment structure and methods of use thereof |
WO2015138343A1 (en) | 2014-03-10 | 2015-09-17 | Click Diagnostics, Inc. | Cartridge-based thermocycler |
CN107429281B (en) | 2014-12-31 | 2022-05-27 | 维斯比医学公司 | Apparatus and method for molecular diagnostic testing |
WO2017185067A1 (en) | 2016-04-22 | 2017-10-26 | Click Diagnostics, Inc. | Printed circuit board heater for an amplification module |
WO2017197040A1 (en) | 2016-05-11 | 2017-11-16 | Click Diagnostics, Inc. | Devices and methods for nucleic acid extraction |
USD800331S1 (en) | 2016-06-29 | 2017-10-17 | Click Diagnostics, Inc. | Molecular diagnostic device |
CN110325652A (en) | 2016-06-29 | 2019-10-11 | 易捷仪器诊断股份有限公司 | Use the device and method of flow cell detection molecules |
USD800914S1 (en) | 2016-06-30 | 2017-10-24 | Click Diagnostics, Inc. | Status indicator for molecular diagnostic device |
USD800913S1 (en) | 2016-06-30 | 2017-10-24 | Click Diagnostics, Inc. | Detection window for molecular diagnostic device |
US11162130B2 (en) | 2017-11-09 | 2021-11-02 | Visby Medical, Inc. | Portable molecular diagnostic device and methods for the detection of target viruses |
WO2020136234A1 (en) * | 2018-12-28 | 2020-07-02 | Astraveus | Device and method for handling a particle suspension |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998009728A1 (en) * | 1996-09-06 | 1998-03-12 | Central Research Laboratories Limited | Apparatus for, and method of, thermally cycling a sample |
WO1998016313A1 (en) * | 1996-10-12 | 1998-04-23 | Central Research Laboratories Limited | Heating apparatus |
WO2000069560A1 (en) * | 1999-05-14 | 2000-11-23 | Gamera Bioscience Corporation | A centripetally-motivated microfluidics system for performing in vitro hybridization and amplification of nucleic acids |
WO2001007159A2 (en) * | 1999-07-28 | 2001-02-01 | Genset | Integration of biochemical protocols in a continuous flow microfluidic device |
JP2001145486A (en) * | 1999-11-19 | 2001-05-29 | Natl Inst Of Advanced Industrial Science & Technology Meti | Reactor for chemical reaction in micro volume for plurality of specimens |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3507146A (en) * | 1968-02-09 | 1970-04-21 | Webb James E | Method and system for respiration analysis |
CA1335880C (en) * | 1988-07-14 | 1995-06-13 | Thomas P. O'connor | Detection of an antibody and antigen in an immunoassay |
GB8917963D0 (en) * | 1989-08-05 | 1989-09-20 | Scras | Apparatus for repeated automatic execution of a thermal cycle for treatment of biological samples |
US5270183A (en) * | 1991-02-08 | 1993-12-14 | Beckman Research Institute Of The City Of Hope | Device and method for the automated cycling of solutions between two or more temperatures |
US6303343B1 (en) * | 1999-04-06 | 2001-10-16 | Caliper Technologies Corp. | Inefficient fast PCR |
-
2001
- 2001-08-16 AU AUPR7071A patent/AUPR707101A0/en not_active Abandoned
-
2002
- 2002-08-16 AU AU2002322178A patent/AU2002322178B2/en not_active Expired
- 2002-08-16 WO PCT/AU2002/001112 patent/WO2003016558A1/en not_active Application Discontinuation
-
2005
- 2005-06-10 US US11/149,217 patent/US7709250B2/en active Active
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998009728A1 (en) * | 1996-09-06 | 1998-03-12 | Central Research Laboratories Limited | Apparatus for, and method of, thermally cycling a sample |
WO1998016313A1 (en) * | 1996-10-12 | 1998-04-23 | Central Research Laboratories Limited | Heating apparatus |
WO2000069560A1 (en) * | 1999-05-14 | 2000-11-23 | Gamera Bioscience Corporation | A centripetally-motivated microfluidics system for performing in vitro hybridization and amplification of nucleic acids |
WO2001007159A2 (en) * | 1999-07-28 | 2001-02-01 | Genset | Integration of biochemical protocols in a continuous flow microfluidic device |
JP2001145486A (en) * | 1999-11-19 | 2001-05-29 | Natl Inst Of Advanced Industrial Science & Technology Meti | Reactor for chemical reaction in micro volume for plurality of specimens |
Non-Patent Citations (1)
Title |
---|
DATABASE WPI Derwent World Patents Index; Class B04, AN 2001-599654/68 * |
Cited By (92)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
USRE45539E1 (en) | 2003-03-14 | 2015-06-02 | Lawrence Livermore National Security, Llc | Method for chemical amplification based on fluid partitioning in an immiscible liquid |
USRE48788E1 (en) | 2003-03-14 | 2021-10-26 | Lawrence Livermore National Security, Llc | Chemical amplification based on fluid partitioning |
USRE43365E1 (en) | 2003-03-14 | 2012-05-08 | Lawrence Livermore National Security, Llc | Apparatus for chemical amplification based on fluid partitioning in an immiscible liquid |
USRE46322E1 (en) | 2003-03-14 | 2017-02-28 | Lawrence Livermore National Security, Llc | Method for chemical amplification based on fluid partitioning in an immiscible liquid |
USRE41780E1 (en) | 2003-03-14 | 2010-09-28 | Lawrence Livermore National Security, Llc | Chemical amplification based on fluid partitioning in an immiscible liquid |
USRE47080E1 (en) | 2003-03-14 | 2018-10-09 | Lawrence Livermore National Security, Llc | Chemical amplification based on fluid partitioning |
US7622076B2 (en) | 2003-09-05 | 2009-11-24 | Stokes Bio Limited | Microfluidic analysis system |
US11807902B2 (en) | 2003-09-05 | 2023-11-07 | Stokes Bio Ltd. | Microfluidic analysis system |
WO2005023427A1 (en) * | 2003-09-05 | 2005-03-17 | Stokes Bio Limited | A microfluidic analysis system |
US20100092987A1 (en) * | 2003-09-05 | 2010-04-15 | Stokes Bio Limited | Microfluidic analysis system |
US20100120635A1 (en) * | 2003-09-05 | 2010-05-13 | Stokes Bio Limited | Sample dispensing |
US8968659B2 (en) * | 2003-09-05 | 2015-03-03 | Stokes Bio Limited | Sample dispensing |
US9597644B2 (en) | 2003-09-05 | 2017-03-21 | Stokes Bio Limited | Methods for culturing and analyzing cells |
US10676786B2 (en) | 2003-09-05 | 2020-06-09 | Stokes Bio Ltd. | Microfluidic analysis system |
US10967338B2 (en) | 2003-09-05 | 2021-04-06 | Stokes Bio Ltd. | Methods of releasing and analyzing cellular components |
US20130040381A1 (en) * | 2004-01-28 | 2013-02-14 | Marshall University Research Corporation | Apparatus and method for a continuous rapid thermal cycle system |
US8986982B2 (en) * | 2004-01-28 | 2015-03-24 | Marshall University Research Corporation | Apparatus and method for a continuous rapid thermal cycle system |
EP1784505A4 (en) * | 2004-09-02 | 2012-12-19 | Bioneer Corp | Miniaturized apparatus for real-time monitoring |
EP1784505A1 (en) * | 2004-09-02 | 2007-05-16 | Bioneer Corporation | Miniaturized apparatus for real-time monitoring |
EP1982195A1 (en) * | 2006-02-02 | 2008-10-22 | Corbett Life Science Pty Ltd | Thermocycler and sample port |
US8124413B2 (en) | 2006-02-02 | 2012-02-28 | Corbett Life Science Pty Ltd | Thermocycler and sample port |
US9352322B2 (en) | 2006-02-02 | 2016-05-31 | Qiagen Instruments Ag | Thermocycler and sample port |
EP1982195A4 (en) * | 2006-02-02 | 2010-07-07 | Corbett Life Science Pty Ltd | Thermocycler and sample port |
WO2007091230A1 (en) * | 2006-02-07 | 2007-08-16 | Stokes Bio Limited | A microfluidic analysis system |
US11772096B2 (en) | 2006-02-07 | 2023-10-03 | Stokes Bio Ltd. | System for processing biological sample |
US10730051B2 (en) | 2006-02-07 | 2020-08-04 | Stokes Bio Ltd. | Liquid bridge and system |
US11084039B2 (en) | 2006-02-07 | 2021-08-10 | Stokes Bio Ltd. | Microfluidic analysis system |
US10741034B2 (en) | 2006-05-19 | 2020-08-11 | Apdn (B.V.I.) Inc. | Security system and method of marking an inventory item and/or person in the vicinity |
US8550503B2 (en) | 2006-09-28 | 2013-10-08 | Stokes Bio Ltd. | Microfluidic connector |
JP2010506187A (en) * | 2006-10-13 | 2010-02-25 | ロディア オペレーションズ | Method and apparatus for determining at least one parameter of physicochemical transformation and / or chemical transformation and corresponding screening method |
WO2008043922A3 (en) * | 2006-10-13 | 2008-06-19 | Rhodia Operations | Method and installation for determining at least one parameter of a physical and/or chemical conversion, and corresponding screening method |
FR2907227A1 (en) * | 2006-10-13 | 2008-04-18 | Rhodia Recherches & Tech | METHOD AND FACILITY FOR DETERMINING AT LEAST ONE PARAMETER OF A PHYSICAL AND / OR CHEMICAL TRANSFORMATION AND CORRESPONDING SCREENING METHOD |
JP2011513743A (en) * | 2008-03-03 | 2011-04-28 | ロディア オペレーションズ | Method and apparatus for determining at least one parameter of physical and / or chemical transition |
WO2009157695A3 (en) * | 2008-06-23 | 2010-03-25 | Bioneer Corporation | Thermal cycling reaction block and continuous real-time monitoring apparatus using the same |
US9205425B2 (en) | 2008-06-23 | 2015-12-08 | Bioneer Corporation | Thermal cycling reaction block and continuous real-time monitoring apparatus using the same |
US10512910B2 (en) | 2008-09-23 | 2019-12-24 | Bio-Rad Laboratories, Inc. | Droplet-based analysis method |
US9636682B2 (en) | 2008-09-23 | 2017-05-02 | Bio-Rad Laboratories, Inc. | System for generating droplets—instruments and cassette |
US9248417B2 (en) | 2008-09-23 | 2016-02-02 | Bio-Rad Laboratories, Inc. | System for droplet-based assays using an array of emulsions |
US9126160B2 (en) | 2008-09-23 | 2015-09-08 | Bio-Rad Laboratories, Inc. | System for forming an array of emulsions |
US10279350B2 (en) | 2008-09-23 | 2019-05-07 | Bio-Rad Laboratories, Inc. | Method of generating droplets |
US9243288B2 (en) | 2008-09-23 | 2016-01-26 | Bio-Rad Laboratories, Inc. | Cartridge with lysis chamber and droplet generator |
US11633739B2 (en) | 2008-09-23 | 2023-04-25 | Bio-Rad Laboratories, Inc. | Droplet-based assay system |
US11612892B2 (en) | 2008-09-23 | 2023-03-28 | Bio-Rad Laboratories, Inc. | Method of performing droplet-based assays |
US9417190B2 (en) | 2008-09-23 | 2016-08-16 | Bio-Rad Laboratories, Inc. | Calibrations and controls for droplet-based assays |
US9492797B2 (en) | 2008-09-23 | 2016-11-15 | Bio-Rad Laboratories, Inc. | System for detection of spaced droplets |
US8633015B2 (en) | 2008-09-23 | 2014-01-21 | Bio-Rad Laboratories, Inc. | Flow-based thermocycling system with thermoelectric cooler |
US9216392B2 (en) | 2008-09-23 | 2015-12-22 | Bio-Rad Laboratories, Inc. | System for forming an array of emulsions |
US10258989B2 (en) | 2008-09-23 | 2019-04-16 | Bio-Rad Laboratories, Inc. | Method of making a device for generating droplets |
US10258988B2 (en) | 2008-09-23 | 2019-04-16 | Bio-Rad Laboratories, Inc. | Device for generating droplets |
US9623384B2 (en) | 2008-09-23 | 2017-04-18 | Bio-Rad Laboratories, Inc. | System for transporting emulsions from an array to a detector |
US9132394B2 (en) | 2008-09-23 | 2015-09-15 | Bio-Rad Laboratories, Inc. | System for detection of spaced droplets |
US9649635B2 (en) | 2008-09-23 | 2017-05-16 | Bio-Rad Laboratories, Inc. | System for generating droplets with push-back to remove oil |
US9764322B2 (en) | 2008-09-23 | 2017-09-19 | Bio-Rad Laboratories, Inc. | System for generating droplets with pressure monitoring |
US11130134B2 (en) | 2008-09-23 | 2021-09-28 | Bio-Rad Laboratories, Inc. | Method of performing droplet-based assays |
US11130128B2 (en) | 2008-09-23 | 2021-09-28 | Bio-Rad Laboratories, Inc. | Detection method for a target nucleic acid |
US9156010B2 (en) | 2008-09-23 | 2015-10-13 | Bio-Rad Laboratories, Inc. | Droplet-based assay system |
US8697011B2 (en) | 2009-05-19 | 2014-04-15 | Stokes Bio Limited | Sampling device with immiscible fluid supply tube in counter-flow arrangement |
US8741660B2 (en) | 2009-05-19 | 2014-06-03 | Stokes Bio Limited | Sampling device |
US9387472B2 (en) | 2009-05-19 | 2016-07-12 | Stokes Bio Limited | Sampling device |
US10166522B2 (en) | 2009-09-02 | 2019-01-01 | Bio-Rad Laboratories, Inc. | System for mixing fluids by coalescence of multiple emulsions |
US10677693B2 (en) | 2009-09-02 | 2020-06-09 | Bio-Rad Laboratories, Inc. | System for mixing fluids by coalescence of multiple emulsions |
US9194861B2 (en) | 2009-09-02 | 2015-11-24 | Bio-Rad Laboratories, Inc. | Method of mixing fluids by coalescence of multiple emulsions |
US11964244B2 (en) | 2009-11-12 | 2024-04-23 | Stokes Bio Limited | Methods of releasing and analyzing cellular components |
US11060136B2 (en) | 2010-03-02 | 2021-07-13 | Bio-Rad Laboratories, Inc. | Emulsion chemistry for encapsulated droplets |
US9598725B2 (en) | 2010-03-02 | 2017-03-21 | Bio-Rad Laboratories, Inc. | Emulsion chemistry for encapsulated droplets |
US11866771B2 (en) | 2010-03-02 | 2024-01-09 | Bio-Rad Laboratories, Inc. | Emulsion chemistry for encapsulated droplets |
US10378048B2 (en) | 2010-03-02 | 2019-08-13 | Bio-Rad Laboratories, Inc. | Emulsion chemistry for encapsulated droplets |
US10272432B2 (en) | 2010-03-25 | 2019-04-30 | Bio-Rad Laboratories, Inc. | Device for generating droplets |
US9393560B2 (en) | 2010-03-25 | 2016-07-19 | Bio-Rad Laboratories, Inc. | Droplet transport system for detection |
US9500664B2 (en) | 2010-03-25 | 2016-11-22 | Bio-Rad Laboratories, Inc. | Droplet generation for droplet-based assays |
US10099219B2 (en) | 2010-03-25 | 2018-10-16 | Bio-Rad Laboratories, Inc. | Device for generating droplets |
US10744506B2 (en) | 2010-03-25 | 2020-08-18 | Bio-Rad Laboratories, Inc. | Device for generating droplets |
EP2553085A2 (en) * | 2010-03-26 | 2013-02-06 | Stokes Bio Limited | Devices, systems, and methods for amplifying nucleic acids |
EP2553085A4 (en) * | 2010-03-26 | 2013-08-28 | Stokes Bio Ltd | Devices, systems, and methods for amplifying nucleic acids |
US9089844B2 (en) | 2010-11-01 | 2015-07-28 | Bio-Rad Laboratories, Inc. | System for forming emulsions |
US9222128B2 (en) | 2011-03-18 | 2015-12-29 | Bio-Rad Laboratories, Inc. | Multiplexed digital assays with combinatorial use of signals |
US10190115B2 (en) | 2011-04-25 | 2019-01-29 | Bio-Rad Laboratories, Inc. | Methods and compositions for nucleic acid analysis |
US9347059B2 (en) | 2011-04-25 | 2016-05-24 | Bio-Rad Laboratories, Inc. | Methods and compositions for nucleic acid analysis |
US10760073B2 (en) | 2011-04-25 | 2020-09-01 | Bio-Rad Laboratories, Inc. | Methods and compositions for nucleic acid analysis |
US11939573B2 (en) | 2011-04-25 | 2024-03-26 | Bio-Rad Laboratories, Inc. | Methods and compositions for nucleic acid analysis |
US9885034B2 (en) | 2011-04-25 | 2018-02-06 | Bio-Rad Laboratories, Inc. | Methods and compositions for nucleic acid analysis |
US8951939B2 (en) | 2011-07-12 | 2015-02-10 | Bio-Rad Laboratories, Inc. | Digital assays with multiplexed detection of two or more targets in the same optical channel |
US9399215B2 (en) | 2012-04-13 | 2016-07-26 | Bio-Rad Laboratories, Inc. | Sample holder with a well having a wicking promoter |
US9963740B2 (en) | 2013-03-07 | 2018-05-08 | APDN (B.V.I.), Inc. | Method and device for marking articles |
US9904734B2 (en) | 2013-10-07 | 2018-02-27 | Apdn (B.V.I.) Inc. | Multimode image and spectral reader |
US10282480B2 (en) | 2013-10-07 | 2019-05-07 | Apdn (B.V.I) | Multimode image and spectral reader |
US10047282B2 (en) | 2014-03-18 | 2018-08-14 | Apdn (B.V.I.) Inc. | Encrypted optical markers for security applications |
US10745825B2 (en) | 2014-03-18 | 2020-08-18 | Apdn (B.V.I.) Inc. | Encrypted optical markers for security applications |
US10519605B2 (en) | 2016-04-11 | 2019-12-31 | APDN (B.V.I.), Inc. | Method of marking cellulosic products |
US10995371B2 (en) | 2016-10-13 | 2021-05-04 | Apdn (B.V.I.) Inc. | Composition and method of DNA marking elastomeric material |
US10920274B2 (en) | 2017-02-21 | 2021-02-16 | Apdn (B.V.I.) Inc. | Nucleic acid coated submicron particles for authentication |
WO2019193104A1 (en) | 2018-04-04 | 2019-10-10 | Altana Ag | Effect pigments based on colored hectorites and coated colored hectorites and manufacture thereof |
Also Published As
Publication number | Publication date |
---|---|
AU2002322178B2 (en) | 2007-11-29 |
US7709250B2 (en) | 2010-05-04 |
AUPR707101A0 (en) | 2001-09-06 |
US20050282206A1 (en) | 2005-12-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US7709250B2 (en) | Continuous flow thermal device | |
AU2002322178A1 (en) | Continuous flow thermal device | |
US10744506B2 (en) | Device for generating droplets | |
US9352322B2 (en) | Thermocycler and sample port | |
US10252261B2 (en) | Handling liquid samples | |
US20080145923A1 (en) | High Throughput Device for Performing Continuous-Flow Reactions | |
US20140272996A1 (en) | Droplet generator with collection tube | |
EP3656475A1 (en) | Rapid thermal cycling for sample analyses and processing | |
CA3210271A1 (en) | Droplet-based assay system | |
US20050158847A1 (en) | Centrifugal array processing device | |
JP2005296017A (en) | Nucleic acid amplification reaction apparatus, chemical chain reaction apparatus, apparatus for simultaneously carrying out nucleic acid amplification reaction including denaturation, annealing and extension process and method for carrying out nucleic acid amplification reaction | |
EP2473618A1 (en) | System for mixing fluids by coalescence of multiple emulsions | |
US20100240048A1 (en) | Biological sample reaction chip, biological sample charging device, biological sample quantifying device, and biological sample reaction method | |
Meldrum et al. | ACAPELLA-1K, a capillary-based submicroliter automated fluid handling system for genome analysis | |
CN113174327A (en) | Stirring control method based on PCR amplification bin | |
JP2010217162A (en) | Biological sample reaction container, biological sample charging device, biological sample quantifying device, and biological sample reaction method |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BY BZ CA CH CN CO CR CU CZ DE DM DZ EC EE ES FI GB GD GE GH HR HU ID IL IN IS JP KE KG KP KR LC LK LR LS LT LU LV MA MD MG MN MW MX MZ NO NZ OM PH PL PT RU SD SE SG SI SK SL TJ TM TN TR TZ UA UG US UZ VC VN YU ZA ZM Kind code of ref document: A1 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ OM PH PL PT RO RU SD SE SG SI SK SL TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): GH GM KE LS MW MZ SD SL SZ UG ZM ZW AM AZ BY KG KZ RU TJ TM AT BE BG CH CY CZ DK EE ES FI FR GB GR IE IT LU MC PT SE SK TR BF BJ CF CG CI GA GN GQ GW ML MR NE SN TD TG Kind code of ref document: A1 Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR IE IT LU MC NL PT SE SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
REG | Reference to national code |
Ref country code: DE Ref legal event code: 8642 |
|
122 | Ep: pct application non-entry in european phase | ||
WWE | Wipo information: entry into national phase |
Ref document number: 2002322178 Country of ref document: AU |
|
WWE | Wipo information: entry into national phase |
Ref document number: 11149217 Country of ref document: US |
|
NENP | Non-entry into the national phase |
Ref country code: JP |
|
WWW | Wipo information: withdrawn in national office |
Country of ref document: JP |
|
ENP | Entry into the national phase |
Ref document number: 2002322178 Country of ref document: AU Date of ref document: 20020816 Kind code of ref document: B |