Library preparation systems and methods

Abstract

A library preparation system and method is disclosed, the apparatus including a plate receptacle for receiving a plate having wells, a magnet, a thermocycler for regulating the temperature of samples in the wells of the plate, and an actuator for moving the magnet relative to the plate receptacle.

Description

[Technical field]

[0001] Related Applications This application claims the benefit of and priority to U.S. Provisional Patent Application No. 63 / 325,049, filed March 29, 2022, the contents of which are incorporated by reference herein in their entirety for all purposes. [Background technology]

[0002] A DNA library can be prepared to allow the sample to be sequenced. Summary of the Invention

[0003] The shortcomings of the prior art can be overcome and advantages achieved through the provision of library preparation systems and methods, as described below in this disclosure. Various implementations of the devices and methods are described below, and the devices and methods, including and excluding the additional implementations listed below, in any combination (provided that such combinations are not inconsistent), can overcome these shortcomings and achieve the benefits described herein.

[0004] In a first implementation, the apparatus includes a plate receptacle, a magnet, a thermocycler, and an actuator. The plate receptacle includes a thermal block and an insert. The thermocycler is in thermal communication with the plate receptacle. The plate receptacle is for receiving a plate having wells, and the thermocycler is for regulating a temperature of a sample in the wells of the plate. The actuator is for moving the magnet relative to the plate receptacle, and the insert is operable to transfer a magnetic field from the magnet to the plate receptacle.

[0005] In a second implementation, the method includes adjusting the temperature of samples in wells of a plate disposed in a plate receptacle including a thermal block and an insert using a thermocycler, dispensing beads into the wells of the plate and dispensing a first reagent into the wells of the plate with the plate disposed in the plate receptacle, moving a magnet toward the insert to allow the insert to transfer a magnetic field from the magnet to the plate receptacle and attract the beads toward the magnet, and aspirating the first reagent from the wells with the plate disposed in the plate receptacle.

[0006] In a third implementation, an apparatus includes a plate receptacle, a magnet, a thermocycler, and an actuator, where the plate receptacle is for receiving a plate having wells, the thermocycler is for regulating a temperature of samples in the wells of the plate, and the actuator is for moving the magnet relative to the plate receptacle.

[0007] Furthermore, in accordance with the first, second, and / or third implementations described above, the apparatus and / or method may further include or include any one or more of the following:

[0008] In an implementation, the plate receptacle includes a well receptacle and the insert is positioned within the thermal block.

[0009] In another implementation, the insert at least partially defines the well receptacle.

[0010] In another implementation, the insert includes at least one of a ferrous material, nickel, mu metal, or a cobalt alloy.

[0011] In another implementation, the insert extends horizontally through the thermal block.

[0012] In another implementation, the apparatus includes a heat sink coupled to the thermocycler.

[0013] In another implementation, the heat sink includes a liquid cooled heat sink.

[0014] In another implementation, the apparatus includes opposing first and second brackets positioned on opposite sides of the plate receptacle, the magnets include a first magnet coupled to the first bracket and a second magnet coupled to the second bracket, and the actuators include a first actuator that moves the first bracket and a second actuator that moves the second bracket relative to the plate receptacle to enable the first magnet and the second magnet to act on the plate receptacle.

[0015] In another implementation, the thermocycler includes a first thermoelectric cooler positioned on a first side of the plate receptacle and a second thermoelectric cooler positioned on a second side of the plate receptacle.

[0016] In another implementation, a heat sink is coupled to each of the first thermoelectric cooler and the second thermoelectric cooler.

[0017] In another implementation, the magnet includes a switchable magnet.

[0018] In another implementation, the actuator is for actuating a switchable magnet.

[0019] In another implementation, the plate receptacle includes a plurality of well receptacles and the insert includes a plurality of cup-shaped pins, each cup-shaped pin positioned beneath one of the well receptacles.

[0020] In another implementation, the apparatus includes a U-shaped bracket carrying a magnet and having an end, and the insert includes a magnetically permeable material extending from a thermal block. The thermal block and the magnetically permeable material define a well receptacle.

[0021] In another implementation, the magnetically permeable material includes at least one of a ferrous material, nickel, mu metal, or a cobalt alloy.

[0022] In another implementation, the thermocycler is positioned within a U-shaped bracket.

[0023] In another implementation, the magnet includes a first magnet and a second magnet. The apparatus includes a first bracket carrying the first magnet and having a first and second end, and a second bracket carrying the second magnet and having a first and second end. The first bracket and the second bracket face each other. The insert includes a first magnetically permeable material extending from the thermal block and a second magnetically permeable material extending from the thermal block. The thermal block, the first magnetically permeable material, and the second magnetically permeable material define a well receptacle.

[0024] In another implementation, the actuator moves the first bracket and the second bracket to allow a first magnetically permeable material to be coupled to and extend between first ends of the first bracket and the second bracket and to allow a second magnetically permeable material to be coupled to and extend between second ends of the first bracket and the second bracket.

[0025] In another implementation, the magnet includes a first magnet and a second magnet. The apparatus includes a first U-shaped bracket carrying the first magnet and having an end, and a second U-shaped bracket carrying the second magnet and having an end. The insert includes a first magnetically permeable material extending from the thermal block and a second magnetically permeable material extending from the thermal block. The thermal block, the first magnetically permeable material, and the second magnetically permeable material define a well receptacle.

[0026] In another implementation, the actuator includes a first actuator and a second actuator, the first actuator for moving the first U-shaped bracket to allow a first magnetically permeable material to be coupled to and extend between ends of the first U-shaped bracket, and the second actuator for moving the second U-shaped bracket to allow a second magnetically permeable material to be coupled to and extend between ends of the second U-shaped bracket.

[0027] In another implementation, the apparatus includes a stage including a thermocycler including a thermal block defining a first well receptacle, and the magnet includes a second well receptacle spaced apart from the first well receptacle.

[0028] In another implementation, the magnet includes a plurality of ring magnets, each ring magnet surrounding one of the second well receptacles.

[0029] In another implementation, the method includes performing a series of temperature adjustments using a thermocycler.

[0030] In another implementation, the method includes moving the magnet away from the insert.

[0031] In another implementation, the method includes dispensing a second reagent into wells of the first plate.

[0032] In another implementation, the method includes moving a magnet toward the insert to allow the insert to transfer a magnetic field from the magnet to the plate receptacle and attract the beads toward the magnet.

[0033] In another implementation, the method includes aspirating a second reagent and the sample from the wells of the plate.

[0034] In another implementation, the method includes dispensing a second reagent and the sample into wells of a second plate.

[0035] In another implementation, the thermocycler is for performing a series of temperature adjustments.

[0036] In another implementation, the beads are for dispensing into wells of a plate and the first reagent is for dispensing into the wells of the plate.

[0037] In another implementation, the actuator is for moving a magnet towards the plate receptacle to attract the beads towards the magnet and the first reagent is for being aspirated from the well.

[0038] In another implementation, the second reagent is for dispensing into the wells of the first plate.

[0039] In another implementation, the actuator is for moving a magnet toward a plate receptacle to attract beads toward the magnet, a second reagent and sample are aspirated from the wells of the plate, and a second reagent and sample are dispensed into the wells of a second plate.

[0040] In another implementation, the thermocycler is positioned below the plate receptacle.

[0041] In another implementation, the plate receptacle has a thermal block that defines a well receptacle, and the thermocycler is positioned below the well receptacle.

[0042] In another implementation, the apparatus includes a heat sink coupled to the thermocycler.

[0043] In another implementation, the apparatus includes opposing first and second brackets positioned on opposite sides of the plate receptacle, the magnets have a first magnet coupled to the first bracket and a second magnet coupled to the second bracket, and the actuators include a first actuator that moves the first bracket and a second actuator that moves the second bracket relative to the plate receptacle to enable the first magnet and the second magnet to act on the plate receptacle.

[0044] In another implementation, the thermocycler has a first thermoelectric cooler positioned on a first side of the plate receptacle and a second thermoelectric cooler positioned on a second side of the plate receptacle.

[0045] In another implementation, an apparatus includes a heat sink coupled to each of the first thermoelectric cooler and the second thermoelectric cooler.

[0046] In another implementation, the plate receptacle includes a thermal block and a ferrous material insert within the thermal block.

[0047] In another implementation, the actuator moves the bracket and magnet relative to the plate receptacle, allowing the magnet to act on the plate receptacle.

[0048] In another implementation, the magnet comprises a switchable magnet.

[0049] In another implementation, the actuator is for actuating a switchable magnet.

[0050] In another implementation, the plate receptacle has a plurality of well receptacles and a plurality of cup-shaped pins, each cup-shaped pin positioned beneath one of the well receptacles.

[0051] In another implementation, the apparatus includes a U-shaped bracket carrying a magnet and having an end, the plate receptacle having a thermal block and a magnetically permeable material extending from the thermal block, the thermal block and the magnetically permeable material defining a well receptacle.

[0052] In another implementation, the actuator moves a U-shaped bracket relative to the magnetically permeable material to allow the magnet to act on the magnetically permeable material.

[0053] In another implementation, the thermocycler is positioned within a U-shaped bracket.

[0054] In another implementation, the magnetically permeable material includes mu-metal.

[0055] In another implementation, the magnet includes a first magnet and a second magnet. The apparatus also includes a first bracket carrying the first magnet and having a first and second end, and a second bracket carrying the second magnet and having a first and second end. The first bracket and the second bracket face each other. The plate receptacle includes a thermal block, a first magnetically permeable material extending from the thermal block, and a second magnetically permeable material extending from the thermal block. The thermal block, the first magnetically permeable material, and the second magnetically permeable material define a well receptacle.

[0056] In another implementation, the actuator moves the first bracket and the second bracket to allow a first magnetically permeable material to be coupled to and extend between first ends of the first bracket and the second bracket and to allow a second magnetically permeable material to be coupled to and extend between second ends of the first bracket and the second bracket.

[0057] In another implementation, the thermocycler is positioned between the first bracket and the second bracket.

[0058] In another implementation, the magnet has a first magnet and a second magnet. The apparatus also includes a first U-shaped bracket carrying the first magnet and having an end, and a second U-shaped bracket carrying the second magnet and having an end. The plate receptacle has a thermal block, a first magnetically permeable material extending from the thermal block, and a second magnetically permeable material extending from the thermal block. The thermal block, the first magnetically permeable material, and the second magnetically permeable material define a well receptacle.

[0059] In another implementation, the actuator includes a first actuator and a second actuator, the first actuator for moving the first U-shaped bracket to allow a first magnetically permeable material to be coupled to and extend between ends of the first U-shaped bracket, and the second actuator for moving the second U-shaped bracket to allow a second magnetically permeable material to be coupled to and extend between ends of the second U-shaped bracket.

[0060] In another implementation, the thermocycler is positioned within the second U-shaped bracket, the stage includes a thermocycler having a thermal block defining a first well receptacle, and the magnet includes a second well receptacle spaced apart from the first well receptacle.

[0061] In another implementation, the magnet includes a plurality of ring magnets, each ring magnet surrounding one of the second well receptacles.

[0062] In another implementation, the stage is for aligning the first well receptacle or the second well receptacle with the second well receptacle.

[0063] It is understood that all combinations of the foregoing and additional concepts, described in more detail below (provided such concepts are not mutually inconsistent), are considered to be part of the subject matter disclosed herein and / or may be combined to achieve particular benefits of particular embodiments described herein. In particular, all combinations of claimed subject matter appearing at the end of this disclosure are considered to be part of the subject matter disclosed herein. [Brief description of the drawings]

[0064] [Figure 1] 1 shows a schematic diagram of an implementation of a system according to the teachings of the present disclosure. [Diagram 2] A cross-sectional view of an implementation of a plate receptacle, a thermocycler, a pair of magnets, and a pair of actuators, with the magnet in a first position, that may be used to implement the plate receptacle, thermocycler, magnet, and actuator of Figure 1. [Diagram 3] 3 is another cross-sectional view of the plate receptacle, thermocycler, pair of magnets, and pair of actuators of FIG. 2, with the magnets in a second position. [Figure 4] 2 is a cross-sectional view of an implementation of a plate receptacle, thermocycler, magnet, and actuator, with the magnet in a first position, that may be used to implement the plate receptacle, thermocycler, magnet, and actuator of FIG. 1. [Diagram 5] 5 is a cross-sectional view of the plate receptacle, thermocycler, magnet, and actuator of FIG. 4, with the magnet in a second position. [Figure 6] 2 is a cross-sectional view of an implementation of a plate receptacle, thermocycler, magnet, and actuator, with the magnet in a first position, that may be used to implement the plate receptacle, thermocycler, magnet, and actuator of FIG. 1. [Figure 7] 7 is a cross-sectional view of the plate receptacle, thermocycler, magnet, and actuator of FIG. 6, with the magnet in a second position. [Figure 8]2 is a cross-sectional view of an implementation of a plate receptacle, thermocycler, magnet, and actuator that can be used to implement the plate receptacle, thermocycler, magnet, and actuator of FIG. 1. [Figure 9] 2 is a cross-sectional view of an implementation of a plate receptacle, a thermocycler, a first magnet, a second magnet, and an actuator that can be used to implement the plate receptacle, thermocycler, magnet, and actuator of FIG. 1. [Figure 10] A cross-sectional view of an implementation of a plate receptacle, a thermocycler, a first magnet, a second magnet, a first actuator, and a second actuator that can be used to implement the plate receptacle, thermocycler, magnet, and actuator of Figure 1. [Figure 11] A cross-sectional view of an implementation of another plate receptacle, thermocycler, first magnet, second magnet, first actuator, and second actuator that can be used to implement the first plate receptacle, thermocycler, magnet, and actuator of Figure 1. [Figure 12] A cross-sectional view of a stage including a thermocycler and magnet that can be used to implement the thermocycler and magnet of Figure 1 and / or that can be used to implement the system of Figure 1, as well as an implementation of another stage. [Figure 13] FIG. 12 is a cross-sectional view of the stage of FIG. 11 showing the wells of the plate positioned in the second well receptacle. [Figure 14] 2 is a cross-sectional view of an implementation of a stage and coolant system that can be used to implement the thermocycler and magnet of FIG. 1 and / or that can be used to implement the system of FIG. 1. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0065] Although the following text discloses a detailed description of implementations of methods, apparatus, and / or products, it should be understood that the legal scope of ownership is defined by the claims at the end of this patent. Therefore, the following "Description of the Preferred Embodiments" should be construed as examples only and does not describe all possible implementations, as describing all possible implementations would be impractical, if not impossible. Numerous alternative implementations may be implemented using either current technology or technology developed after the filing date of this patent. It is contemplated that such alternative implementations would still fall within the scope of the claims.

[0066] 1 shows a schematic diagram of an implementation of a system 100 according to the teachings of the present disclosure. The system 100 can be used to perform amplification and clean-up operations, for example, when preparing a DNA library for sequencing applications. Thus, the system 100 can perform a DNA library preparation workflow including, for example, an amplification process, a clean-up process.

[0067] The system 100 is shown carrying plates 120, 121 having wells 122 and, in the implementation shown, includes a working area 106 having a magnet 150, a thermocycler 152, a first plate receptacle 158, a second plate receptacle 160, and an actuator 166. The magnet 150 may be a permanent magnet or an electromagnet. The plate receptacles 158, 160 may be referred to as plate stations. The different wells 122 of the first plate 120 may contain different samples 124. The samples 124 may be biological samples of human, animal, plant, bacterial, or fungal origin. Other sources of obtaining biological samples may prove suitable.

[0068] An actuator 166 moves the magnet 150 relative to the first plate receptacle 158 during operation. The actuator 166 can move the magnet 150 between an upper position, in which the magnet 150 influences any plates positioned on the first plate receptacle 158, and a lower position, in which the magnet 150 does not influence any plates positioned on the first plate receptacle 158. This may not work well for Figures 6 and 7, but may be applicable to the rest.

[0069] The magnet 150 being moved relative to the first plate receptacle 158 and any plates 120, 121 positioned thereon allows less area on the working area 106 to be consumed. The magnet 150 can also be moved with a relatively high degree of reliability compared to the alternative of moving one of the sample-loaded plates 120, 121 to a separate magnet station. The magnet 150 can be implemented in a 9econd9 array configuration to strengthen and focus the corresponding magnetic field.

[0070] The reagents 168 and / or samples 124 are dispensed into the wells 122 of the first plate 120, and the lid 170 is positioned on the first plate 120 such that the lid 170 covers the wells 122 of the first plate 120. Alternatively, the lid 170 may be movably coupled to the thermocycler 152 (see, e.g., FIG. 6). The lid 170 need not be disposable in such implementations.

[0071] The thermocycler 152 is aligned with and / or in thermal communication with the first plate receptacle 158, and the thermocycler 152 regulates the temperature of the samples 124 in the wells 122 of the first plate 120. The enzymes and reagents 168 amplify the nucleic acids in the samples 124.

[0072] The thermocycler 152 can be programmed to perform a series of temperature adjustments that allow the enzymes and reagents 168 in the wells 122 to amplify the nucleic acids of the sample 124 in the same wells 122. Thus, the thermocycler 152 and / or the magnet 150 can act on the plates 120, 121 received in the first plate receptacle 158. Alternatively, the thermocycler 152 can be spaced apart from the magnet 150.

[0073] The system 100 can perform a clean-up process after the amplification process is performed. The beads 172 can be dispensed into the wells 140 of the first plate 120 as part of the clean-up process, and a first reagent can be dispensed into the wells 122 of the first plate 120. The first reagent can be a bead buffer, and the sample 124 can bind to the beads 172 in the presence of the bead buffer.

[0074] The actuator 166 moves the magnet 150 towards the first plate receptacle 158, which attracts the beads 172 towards the magnet 150. The beads 172 and the samples 124 bound to the beads 172 can be positioned towards the bottom of the wells 122 of the first plate 120 or on the sides of the wells 122. The actuator 166 can change the position of the magnet 150 relative to the first plate receptacle 158 as a way of adjusting the magnetic field strength acting on the beads 172. When the beads 172 are on the sides of the wells 122, the tip of the pipette can easily access the wells 122. However, the magnet 150 can cause the beads 172 to be present anywhere within the wells 122.

[0075] A first reagent may be aspirated from the wells 122 of the first plate 120 and dispensed into the wells 122 of the first plate 120. A second reagent may be dispensed into the wells 122 of the first plate 120. The second reagent may be an elution buffer that releases the sample 124 from binding to the beads 172, and in particular, releases DNA associated with the sample 124 from binding to the beads 172. The actuator 166 moves the magnet 150 towards the first plate receptacle 158, attracting the beads 172 towards the magnet 150, thus suspending the second reagent and the sample 124 within the wells 122.

[0076] The second reagent and sample 124 may be aspirated from the well 122 of the first plate 120, for example, using a pipette tip, and the second reagent and sample 124 may be dispensed into the well 128 of the second plate 121.

[0077] 2 is a cross-sectional view of an implementation of a plate receptacle 800, a thermocycler 802, a pair of magnets 804, 806, and a pair of actuators 808, 810 with the magnets 804, 806 in a first position, which may be used to implement the plate receptacle 158, thermocycler 152, magnet 150, and actuator 166 of FIG. 1. The plate receptacle 800 has a thermal block 811 that defines a well receptacle 812, and the thermocycler 802 is positioned below the well receptacle 812 and therefore below the plate receptacle 800. The thermal block 811 may be a metal and / or a 3D heat pipe. The metal may include aluminum, silver, copper, and / or brass. The thermocycler 802 may additionally or alternatively be a resistive heater.

[0078] An insert 814 is shown positioned within the thermal block 811 and at least partially defining a well receptacle 812, and a heat sink 815 is coupled to the thermocycler 802. The insert 814 may be a ferrous material insert and / or the insert may be operable to transfer a magnetic field from the magnets 804, 806 to the plate receptacle 800. The insert 814 extends horizontally through the thermal block 811 in the implementation shown. The thermal block 811 may be made of a metal such as aluminum or silver, and the insert 814 may be made of iron. A well 816 of a plate 819 is shown received by the well receptacle 812. The plate 819 may be used to implement any one of the first plate 120 or the second plate 121 of FIG.

[0079] The actuator assembly 820 is shown having opposing first and second brackets 822, 824 positioned on either side of the plate receptacle 800 in the illustrated implementation. The first magnet 804 and first actuator 808 are coupled to the first bracket 822, and the second magnet 806 and second actuator 810 are coupled to the second bracket 824. Although the actuator assembly 820 is shown to include first and second actuators 808, 810, alternatively, one actuator may be provided. The first actuator 808 moves the first bracket 822 and the second actuator 810 moves the second bracket 824 in a direction generally indicated by arrow 826 relative to the plate receptacle 800 during operation to allow the first magnet 804 and second magnet 806 to act on the plate 819. The magnets 804, 806 are shown in Figure 2 spaced apart from the plate 819 and the insert 814 such that the beads 172 are suspended within the wells 816. The first position of the actuator assembly 820 shown in Figure 2 may be referred to as the off position, and the second position of the actuator assembly 820 shown in Figure 3 may be referred to as the on position.

[0080] 3 is another cross-sectional view of the plate receptacle 800, thermocycler 802, pair of magnets 804, 806, and pair of actuators 808, 810 of FIG. 2 with the magnets 804, 806 in a second position. The magnets 804, 806 are shown adjacent to the insert 814, such that the magnets 804, 806 magnetize the insert 814 and the beads 172 are attracted towards the corresponding magnets 804, 806. The beads 172 are attracted towards the side of the well 816 in the implementation shown. The actuators 808, 810 can change the position of the magnets 804, 806 relative to the plate receptacle 800 as a way of adjusting the magnetic field strength acting on the beads 172.

[0081] 4 is a cross-sectional view of an implementation of a plate receptacle 900, thermocycler 802, magnet 902, and actuator 904 with magnet 902 in a first position that may be used to implement the first plate receptacle 158, thermocycler 152, magnet 150, and actuator 166 of FIG. 1. The plate receptacle 900 has a thermal block 811 that defines a well receptacle 812, with an insert 905 positioned within the thermal block 811. The insert 905 may be a ferrous material insert. The insert 905 at least partially defines the well receptacle 812 and is shown to be U-shaped.

[0082] 4 has a first thermoelectric cooler 906 positioned on a first side 908 of a plate receptacle 900 and a second thermoelectric cooler 910 positioned on a second side 912 of the plate receptacle 900. A heat sink 815 is coupled to each of the first thermoelectric cooler 906 and the second thermoelectric cooler 910.

[0083] The actuator assembly 914, in the illustrated implementation, is shown with a bracket 916 positioned below the plate receptacle 900. The bracket 916 carries a magnet 902 and is shown to be a U-shaped bracket. The actuator 904 moves the bracket 916 and magnet 902 relative to the plate receptacle 900 in a direction generally indicated by arrow 918, allowing the magnet 902 to act on the plate 819. The magnet 902 is shown spaced apart from the plate 819 and insert 905 in FIG. 4 such that the beads 172 are suspended within the well 816.

[0084] 5 is a cross-sectional view of the plate receptacle 900, thermocycler 802, magnet 902, and actuator 904 of FIG. 4 with the magnet 902 in a second position. The magnet 902 is shown adjacent to the insert 905 such that the magnet 902 magnetizes the insert 905 and the beads 172 are attracted toward the actuator assembly 914 and / or the magnet 902. The insert 905 is U-shaped and has ends 920, 922, and the bracket 916 is also U-shaped and has ends 924, 926 positioned adjacent to the ends 920, 922 of the insert 905, allowing the magnet 902 to magnetize the insert 905 and thus attract the beads 172 toward the insert 905. The bracket 916 is shown with first and second arms 928, 930, and the magnet 902 is shown coupled to the second arm 930. The actuator assembly 914 may alternatively include two of the magnets 902 , and each of the first and second arms 928 , 930 of the bracket 916 may carry one of the magnets 902 .

[0085] 6 is a cross-sectional view of an implementation of a plate receptacle 1000, thermocycler 802, magnet 1002, and actuator 1004 with magnet 1002 in a first position that may be used to implement the first plate receptacle 158, thermocycler 152, magnet 150, and actuator 166 of FIG. 1. The plate receptacle 1000 has a thermal block 811 that defines a well receptacle 812, and inserts 1006, 1008 positioned within the thermal block 811. The inserts 1006, 1008 are positioned below the well receptacle 812 and each have a cup-shaped surface 1010 that faces the corresponding well receptacle 812. The inserts 1006, 1008 may be referred to as cup-shaped pins.

[0086] A portion 1012 of the thermal block 811 is positioned between each of the inserts 1006, 1008 and the corresponding well receptacle 812 for heat transfer. The portion 1012 may be approximately 1 mm thick. A lid 1014 is positioned over the top of the well receptacle 812. The lid 1014 may be operatively coupled to the thermal block 811 and / or may be hingedly coupled to the thermal block 811. The lid 1014 may be a temperature controlled cover that reduces evaporation and improves thermal uniformity. The lid 1014 may be used to implement the lid 170 of FIG. 1.

[0087] The magnet 1002, in the illustrated implementation, is a switchable magnet 1016, and the actuator 1004 actuates the switchable magnet 1016 between a non-magnetized position (first position) shown in Figure 6 and a magnetized position (second position) shown in Figure 7. Thus, the beads 172 are suspended within the well 816 of Figure 6.

[0088] Figure 7 is a cross-sectional view of the plate receptacle 1000, thermocycler 802, magnet 1002, and actuator 1004 of Figure 6, with the magnet 1002 in a second position. The beads 172 are attracted towards the inserts 1006, 1008 and / or switchable magnet 1016 of Figure 7.

[0089] 8 is a cross-sectional view of an implementation of a plate receptacle 1100, a thermocycler 802, a magnet 1102, and an actuator 1104 that may be used to implement the first plate receptacle 158, the thermocycler 152, the magnet 150, and the actuator 166 of FIG. 1. The plate receptacle 1100 has a thermal block 811 that defines a well receptacle 812 and a magnetically permeable material 1106 extending from the thermal block 811. The magnetically permeable material 1106 is shown to at least partially define the well receptacle 812. The magnetically permeable material 1106 extends horizontally through the thermal block 811 in the implementation shown. The magnetically permeable material 1106 may include mu metal, ferritic stainless steel, and / or Metglas.

[0090] The actuator assembly 1108 is shown having a bracket 1110 by which the magnet 1102 is carried. The bracket 1110 is shown as a U-shaped bracket 1112 by which the thermocycler 802 is positioned. The actuator 1104 is coupled to the bracket 1110 and, during operation, moves the bracket 1110 between directions generally indicated by arrows 1114 to enable the magnet 1102 to act on the magnetically permeable material 1106 and attract the beads 172 towards the magnetically permeable material 1106.

[0091] In the implementation shown, the plate 819 has two rows of wells 816, such that the well receptacles 812 and corresponding wells 816 are spaced and / or positioned to allow side access to each of the wells 816. The magnetically permeable material 1106 may route and / or shape the magnetic field as a result of the location of the wells 816. The plate 819 could include additional rows of wells 816, and the magnetically permeable material could still route and / or shape the magnetic field if, for example, the wells 816 were spaced a threshold distance apart.

[0092] 9 is a cross-sectional view of an implementation of a plate receptacle 1200, a thermocycler 802, a first magnet 1202, a second magnet 1204, and an actuator 1206 that may be used to implement the first plate receptacle 158, the thermocycler 152, the magnet 150, and the actuator 166 of FIG. 1. The plate receptacle 1200 has a thermal block 811 defining a well receptacle 812, a first magnetically permeable material 1208 extending from the thermal block 811, and a second magnetically permeable material 1210 extending from the thermal block 811. The first magnetically permeable material 1208 and / or the second magnetically permeable material 1210 may be made of a material in which a magnetic field can be induced when exposed to an external magnetic field. For example, the first magnetically permeable material 1208 / / or the second magnetically permeable material 1210 may include at least one of a ferrous material, nickel, mu metal, or a cobalt alloy.

[0093] The thermal block 811, the first magnetically permeable material 1208, and the second magnetically permeable material 1210 are shown to at least partially define a well receptacle 812. The first magnetically permeable material 1208 and the second magnetically permeable material 1210 extend horizontally through the thermal block 811 and are spaced apart in the implementation shown. The magnetically permeable materials 1208, 1210 may be configured to minimize or reduce the time to focus the beads 141, achieve a threshold spatial distribution of the beads 141 to minimize or reduce any bead loss in the wash operation, and minimize or reduce the liquid volume used in the elution step.

[0094] The actuator assembly 1212 is shown including a first bracket 1214 carrying a first magnet 1202 and having first and second ends 1216, 1218, and a second bracket 1220 carrying a second magnet 1204 and having first and second ends 1222, 1224. The first bracket 1214 and the second bracket 1220 face each other. The thermocycler 802 is positioned between the first bracket 1214 and the second bracket 1220. During operation, the actuator 1206 moves the first bracket 1214 and the second bracket 1220 relative to the first magnetically permeable material 1208 and the second magnetically permeable material 1210 to allow the magnets 1202, 1204 to act on the first magnetically permeable material 1208 and the magnets 1202, 1204 to act on the second magnetically permeable material 1210. The magnets 1202, 1204 acting on the first magnetically permeable material 1208 and the second magnetically permeable material 1210 being spaced apart may generate a magnetic field distribution that draws the beads 172 towards the bottom of the well 816.

[0095] Figure 10 is a cross-sectional view of an implementation of a plate receptacle 1200, a thermocycler 802, a first magnet 1300, a second magnet 1302, a first actuator 1304, and a second actuator 1306 that may be used to implement the first plate receptacle 158, the thermocycler 152, the magnet 150, and the actuator 166 of Figure 1. The plate receptacle 1200 of Figure 9 is substantially the same as the plate receptacle 1200 of Figure 10.

[0096] However, it includes an actuator assembly 1308 that is different from the actuator assembly 1212 of Figure 8. The actuator assembly 1308 of Figure 9 is shown including a first U-shaped bracket 1310 carrying a first magnet 1300 and having ends 1312, 1314, and a second U-shaped bracket 1316 carrying a second magnet 1302 and having ends 1317, 1318. The thermocycler 802 is positioned within the second U-shaped bracket 1316.

[0097] Ends 1312, 1314 of a first U-shaped bracket 1310 are shown positioned to magnetize the first magnetically permeable material 1208, and ends 1317, 1318 of a second U-shaped bracket 1316 are shown positioned to magnetize the second magnetically permeable material 1210. A first actuator 1304 moves the first U-shaped bracket 1310 to allow the first magnetically permeable material 1208 to be coupled to and extend between the ends 1312, 1314 of the first U-shaped bracket 1310, and a second actuator 1306 moves the second U-shaped bracket 1316 to allow the second magnetically permeable material 1210 to be coupled to and extend between the ends 1317, 1318 of the second U-shaped bracket 1316. The first actuator 1304 can move the first U-shaped bracket 1310 relative to the second U-shaped bracket 1316 in a direction generally indicated by arrow 1319, and the second actuator 1306 can move the second U-shaped bracket 1316 relative to the first U-shaped bracket 1310 in a direction generally indicated by arrow 1320. Thus, the first actuator 1304 and the second actuator 1306 are independently actuatable.

[0098] The first U-shaped bracket 1310 can engage and be magnetized with the first magnetically permeable material 1208 during a first operation to capture the bead 172 around a first location 1322 in the well receptacle 812, and the second U-shaped bracket 1316 can engage and be magnetized with the second magnetically permeable material 1210 during a second operation to capture the bead 172 around a second location 1324 in the well receptacle 812. The first operation can be a wash operation and the second operation can be an elution operation.

[0099] 11 is a cross-sectional view of an implementation of another plate receptacle 1250, thermocycler 802, first magnet 1300, second magnet 1302, first actuator 1304, and second actuator 1306 that may be used to implement first plate receptacle 158, thermocycler 152, magnet 150, and actuator 166 of FIG. 1. The plate receptacle 1250 of FIG. 11 is similar to the plate receptacle 1200 of FIG. 9. However, the plate receptacle 1250 of FIG. 12 includes a plating or coating 1252 on a surface 1254 that defines the well receptacle 812. The plating or coating 1252 may reduce exposure of the magnetically permeable material 1208, 1210 to moisture and / or humidity and therefore reduce the likelihood that the magnetically permeable material 1208, 1210 will corrode.

[0100] FIG. 12 is a cross-sectional view of an implementation of a stage 1400 including a thermocycler 1402 and a magnet 1404, as well as another stage 1406, which may be used to implement the thermocycler 152 and magnet 150 of FIG. 1 and / or may be used to implement the system 100 of FIG. 1.

[0101] The thermocycler 1402 includes a thermal block 1408 defining a first well receptacle 1410, and the magnet 1404 has a second well receptacle 1412 spaced apart from the first well receptacle 1410. The magnet 1404 may include ring magnets 1414, each of which surrounds one of the second well receptacles 1412. The stage 1406 positions and / or holds a plate 819 above the stage 1400 during operation, which aligns the first well receptacle 1410 or the second well receptacle 1412 with the plate 819. Stage 1400 positions wells 816 of plate 819 in first well receptacle 1410 or 16th well receptacle 1412, depending on the operation being performed. Stage 1406 can move in a direction generally indicated by arrow 1416, and stage 1400 can move in a direction generally indicated by arrow 1418.

[0102] Figure 13 is a cross-sectional view of the stage 1400, 1406 of Figure 12 showing a well 816 of a plate 819 positioned within a second well receptacle 1412. The magnet 1404 can then act on the well 816 and / or the contents of the well 816.

[0103] Figure 14 is a cross-sectional view of an implementation of a stage 1500 and a coolant system 1502 that may be used to implement the thermocycler 152 and magnet 150 of Figure 1 and / or that may be used to implement the system 100 of Figure 1. The stage 1500 of Figure 14 is similar to the stage 1400 of Figure 12, but includes a heat sink 815 implemented by a liquid-cooled heat sink 1504.

[0104] The coolant system 1502 is shown including fluid lines 1506, a pump 1508, a reservoir 1510 containing a coolant 1511, and a heat exchanger 1512. A heat sink 1504 may be considered part of the coolant system 1502. The fluid lines 1506, in the implementation shown, pass through the heat sink 1504. The heat exchanger 1512 may be referred to as a radiator, and the coolant may be a liquid such as water, antifreeze, and / or ethylene glycol.

[0105] A pump 1508 pumps coolant 1511 from a reservoir 1510 through the fluid lines 1506 and the heat sink 1504 during operation. The coolant 1511 may enter the heat sink 1504 at a first temperature and may draw heat from the thermocycler 152 while in the portion of the fluid lines 1506 in the heat sink 1504, and the coolant 1511 may exit the heat sink 1504 at a second temperature. The second temperature may be higher than the first temperature. The coolant 1511 exiting the heat sink 1504 may enter a heat exchanger 1512. The heating converter 1512 may reduce the temperature of the coolant 1511 from the second temperature back to the first temperature or to an otherwise reduced temperature. The coolant 1511 entering the reservoir 1510 may, for example, be the same or similar in temperature as the coolant 1511 in the reservoir 1510 and / or may not increase the temperature of the coolant 1511 in the reservoir 1510.

[0106] The foregoing description is provided to enable one skilled in the art to practice the various configurations described herein. While the subject technology has been specifically described with reference to various figures and configurations, it should be understood that these are for illustrative purposes only and should not be construed as limiting the scope of the subject technology.

[0107] As used herein, elements or steps described in the singular and followed by the words "a" or "an" should be understood as not excluding a plurality of those elements or steps, unless such exclusion is expressly stated. Moreover, references to "one implementation" are not intended to be interpreted as excluding the existence of additional implementations that also incorporate the recited features. Furthermore, unless expressly stated to the contrary, implementations that "comprising," "including," or "having" an element or elements having a particular characteristic may include additional elements whether or not they have that characteristic. Moreover, the terms "comprising," "including," "having," and the like are used interchangeably herein.

[0108] As used throughout this specification, the terms "substantially," "approximately," and "about" are used to describe and take into account small variations due to processing variations and the like. For example, small variations can refer to ±5% or less, such as ±2% or less, such as ±1% or less, such as ±0.5% or less, such as ±0.2% or less, such as ±0.1% or less, such as ±0.05% or less. In one example, these terms include the situation of no variation, -0%.

[0109] There may be many other ways to implement the subject technology. The various functions and elements described herein may be divided differently than shown without departing from the scope of the subject technology. Various modifications to these implementations may be readily apparent to those skilled in the art, and the general principles defined herein may be applied to other implementations. Thus, many changes and modifications may be made to the subject technology by those skilled in the art without departing from the scope of the subject technology. For example, a different number of a given module or unit may be used, different or multiple types of a given module or unit may be used, a given module or unit may be added, or a given module or unit may be omitted.

[0110] Underlined and / or italicized headings and subheadings are used for convenience only, are not limiting of the subject technology, and are not referred to in connection with interpreting the subject technology description. All structural and functional equivalents to the elements of the various implementations described throughout this disclosure that are known or that later become known to those skilled in the art are expressly incorporated herein by reference and are intended to be encompassed by the subject technology. Furthermore, nothing disclosed herein is intended to be publicly exclusive, regardless of whether such disclosure is expressly set forth in the description above.

[0111] It is to be understood that all combinations of the foregoing concepts and additional concepts, described in more detail below (provided such concepts are not mutually inconsistent), are considered to be part of the subject matter disclosed herein. In particular, all combinations of claimed subject matter appearing at the end of this disclosure are considered to be part of the subject matter disclosed herein.

Claims

1. A plate receptacle including a thermal block and insert, Magnets and A thermocycler that has thermal communication with the aforementioned plate receptacle, Equipped with an actuator, The plate receptacle is for receiving a plate having wells, and the thermocycler is for regulating the temperature of the sample in the wells of the plate. The actuator is for moving the magnet relative to the plate receptacle, The insert is operable to transmit a magnetic field from the magnet to the plate receptacle. Device.

2. The apparatus according to claim 1, wherein the plate receptacle includes a well receptacle, and the insert is positioned within the thermal block.

3. The apparatus according to claim 2, wherein the insert defines the well receptacle at least partially.

4. The apparatus according to claim 1, wherein the insert comprises at least one of iron material, nickel, mu-metal, or cobalt alloy.

5. The apparatus according to claim 1, wherein the insert extends horizontally through the thermal block.

6. The apparatus according to claim 1, further comprising a heat sink coupled to the thermocycler.

7. The apparatus according to claim 6, wherein the heat sink includes a liquid-cooled heat sink.

8. The apparatus according to any one of claims 1 to 7, further comprising opposing first and second brackets positioned on both sides of the plate receptacle, wherein the magnet comprises a first magnet coupled to the first bracket and a second magnet coupled to the second bracket, and the actuator comprises a first actuator for moving the first bracket and a second actuator for moving the second bracket relative to the plate receptacle, thereby enabling the first and second magnets to act on the plate receptacle.

9. The apparatus according to any one of claims 1 to 7, wherein the thermocycler includes a first thermoelectric cooler positioned on a first side of the plate receptacle and a second thermoelectric cooler positioned on a second side of the plate receptacle.

10. The apparatus according to claim 9, further comprising a heat sink coupled to each of the first thermoelectric cooler and the second thermoelectric cooler.

11. The apparatus according to any one of claims 1 to 7, wherein the magnet includes a switchable magnet.

12. The apparatus according to claim 11, wherein the actuator is for operating the switchable magnet.

13. The apparatus according to claim 11, wherein the plate receptacle comprises a plurality of well receptacles, and the insert comprises a plurality of cup-shaped pins, each cup-shaped pin positioned below one of the well receptacles.

14. The apparatus according to any one of claims 1 to 7, further comprising a U-shaped bracket supporting the magnet and having an end, wherein the insert comprises a magnetic permeable material extending from the thermal block, and the thermal block and the magnetic permeable material define a well receptacle.

15. The apparatus according to claim 14, wherein the magnetic permeable material comprises at least one of an iron material, nickel, mu-metal, or a cobalt alloy.

16. The apparatus according to claim 14, wherein the thermocycler is positioned within the U-shaped bracket.

17. The apparatus according to any one of claims 1 to 7, wherein the magnet comprises a first magnet and a second magnet, further comprising a first bracket supporting the first magnet and having first and second ends, and a second bracket supporting the second magnet and having first and second ends, wherein the first bracket and the second bracket face each other, and the insert comprises a first permeable material extending from the thermal block and a second permeable material extending from the thermal block, and the thermal block, the first permeable material and the second permeable material define a well receptacle.

18. The apparatus according to claim 17, wherein the actuator moves the first bracket and the second bracket so that the first magnetic permeable material is coupled to the first ends of the first bracket and the second bracket and extends between them, and the second magnetic permeable material is coupled to the second ends of the first bracket and the second bracket and extends between them.

19. The apparatus according to any one of claims 1 to 7, wherein the magnet comprises a first magnet and a second magnet, further comprising a first U-shaped bracket supporting the first magnet and having an end, and a second U-shaped bracket supporting the second magnet and having an end, and the insert comprises a first permeable material extending from the thermal block and a second permeable material extending from the thermal block, and the thermal block, the first permeable material, and the second permeable material define a well receptacle.

20. The apparatus according to claim 19, wherein the actuator includes a first actuator and a second actuator, the first actuator moving the first U-shaped bracket so that the first permeable material is coupled to the end of the first U-shaped bracket and extends between them, and the second actuator moving the second U-shaped bracket so that the second permeable material is coupled to the end of the second U-shaped bracket and extends between them.

21. The apparatus according to any one of claims 1 to 7, further comprising a stage including the thermocycler, which includes the thermal block defining a first well receptacle, and the magnet, which includes a second well receptacle spaced apart from the first well receptacle.

22. The apparatus according to claim 21, wherein the magnet includes a plurality of ring magnets, each ring magnet surrounding one of the second well receptacles.

23. The temperature of the sample in the wells of a plate placed in a plate receptacle containing a thermal block and insert is controlled using a thermocycler. With the plate placed in the plate receptacle, the beads are dispensed into the wells of the plate, and the first reagent is dispensed into the wells of the plate. The magnet is moved toward the insert, allowing the insert to transmit a magnetic field from the magnet to the plate receptacle, thereby attracting the beads toward the magnet. With the plate placed in the plate receptacle, the first reagent is aspirated from the well. Methods that include...

24. The method according to claim 23, further comprising performing a series of temperature adjustments using the thermocycler.

25. The method according to claim 23, further comprising moving the magnet away from the insert.

26. The method according to any one of claims 23 to 25, further comprising dispensing a second reagent into the wells of a first plate.

27. The method according to claim 26, further comprising moving the magnet toward the insert so that the insert transmits a magnetic field from the magnet to the plate receptacle and attracts the beads toward the magnet.

28. The method according to claim 27, further comprising aspirating the second reagent and the sample from the wells of the plate.

29. The method according to claim 28, further comprising dispensing the second reagent and the sample into the wells of a second plate.

30. Plate receptacles and, Magnets and Thermocycler and, Equipped with an actuator, The plate receptacle is for receiving a plate having wells, the thermocycler is for regulating the temperature of the sample in the wells of the plate, and the actuator is for moving the magnet relative to the plate receptacle. Device.

31. The apparatus according to claim 30, wherein the thermocycler is for performing a series of temperature adjustments.

32. The apparatus according to claim 30, wherein beads are dispensed into the wells of the plate, and the first reagent is dispensed into the wells of the plate.

33. The apparatus according to claim 32, wherein the actuator moves the magnet toward the plate receptacle to attract the beads toward the magnet, and the first reagent is drawn out of the well.

34. The apparatus according to claim 33, wherein the second reagent is dispensed into the wells of the first plate.

35. The apparatus according to claim 34, wherein the actuator moves the magnet toward the plate receptacle to attract the beads toward the magnet, the second reagent and the sample are drawn from the well of the plate, and the second reagent and the sample are dispensed into the well of the second plate.

36. The apparatus according to claim 30, wherein the thermocycler is positioned below the plate receptacle.

37. The apparatus according to claim 30, wherein the plate receptacle includes a thermal block defining a well receptacle, and the thermocycler is positioned below the well receptacle.

38. The apparatus according to claim 30, further comprising a heat sink coupled to the thermocycler.

39. The apparatus according to any one of claims 30 to 38, further comprising opposing first and second brackets positioned on both sides of the plate receptacle, wherein the magnet comprises a first magnet coupled to the first bracket and a second magnet coupled to the second bracket, and the actuator comprises a first actuator for moving the first bracket and a second actuator for moving the second bracket relative to the plate receptacle, thereby enabling the first and second magnets to act on the plate receptacle.

40. The apparatus according to any one of claims 30 to 38, wherein the thermocycler includes a first thermoelectric cooler positioned on a first side of the plate receptacle and a second thermoelectric cooler positioned on a second side of the plate receptacle.

41. The apparatus according to claim 40, further comprising a heat sink coupled to each of the first thermoelectric cooler and the second thermoelectric cooler.

42. The apparatus according to any one of claims 30 to 38, wherein the plate receptacle includes a thermal block and an iron material insert within the thermal block.

43. The apparatus according to any one of claims 30 to 38, wherein the actuator moves the bracket and the magnet relative to the plate receptacle, thereby enabling the magnet to act on the plate receptacle.

44. The apparatus according to any one of claims 30 to 38, wherein the magnet includes a switchable magnet.

45. The apparatus according to claim 44, wherein the actuator is for operating the switchable magnet.

46. The apparatus according to claim 44, wherein the plate receptacle comprises a plurality of well receptacles and a plurality of cup-shaped pins, each cup-shaped pin being positioned below one of the well receptacles.

47. The apparatus according to any one of claims 30 to 38, further comprising a U-shaped bracket supporting the magnet and having an end, wherein the plate receptacle comprises a thermal block and a magnetic permeable material extending from the thermal block, and the thermal block and the magnetic permeable material define a well receptacle.

48. The apparatus according to claim 47, wherein the actuator moves the U-shaped bracket relative to the magnetic permeable material, thereby enabling the magnet to act on the magnetic permeable material.

49. The apparatus according to claim 47, wherein the thermocycler is positioned within the U-shaped bracket.

50. The apparatus according to claim 47, wherein the magnetic permeable material includes a mu-metal.

51. The apparatus according to any one of claims 30 to 38, wherein the magnet comprises a first magnet and a second magnet, further comprising a first bracket supporting the first magnet and having first and second ends, and a second bracket supporting the second magnet and having first and second ends, wherein the first bracket and the second bracket face each other, and the plate receptacle comprises a thermal block, a first permeable material extending from the thermal block, and a second permeable material extending from the thermal block, wherein the thermal block, the first permeable material, and the second permeable material define a well receptacle.

52. The apparatus according to claim 51, wherein the actuator moves the first bracket and the second bracket so that the first permeable material is coupled to the first ends of the first bracket and the second bracket and extends between them, and the second permeable material is coupled to the second ends of the first bracket and the second bracket and extends between them.

53. The apparatus according to claim 51, wherein the thermocycler is positioned between the first bracket and the second bracket.

54. The apparatus according to any one of claims 30 to 38, wherein the magnet comprises a first magnet and a second magnet, further comprising a first U-shaped bracket supporting the first magnet and having an end, and a second U-shaped bracket supporting the second magnet and having an end, and the plate receptacle comprises a thermal block, a first permeable material extending from the thermal block, and a second permeable material extending from the thermal block, and the thermal block, the first permeable material, and the second permeable material define a well receptacle.

55. The apparatus according to claim 54, wherein the actuator includes a first actuator and a second actuator, the first actuator moving the first U-shaped bracket so that the first permeable material is coupled to the end of the first U-shaped bracket and extends between them, and the second actuator moving the second U-shaped bracket so that the second permeable material is coupled to the end of the second U-shaped bracket and extends between them.

56. The apparatus according to claim 53, wherein the thermocycler is positioned within the second U-shaped bracket.

57. The apparatus according to any one of claims 30 to 38, further comprising a stage including a thermocycler including a thermal block defining a first well receptacle, and a magnet including a second well receptacle spaced apart from the first well receptacle.

58. The apparatus according to claim 57, wherein the magnet includes a plurality of ring magnets, each ring magnet surrounding one of the second well receptacles.

59. The apparatus according to claim 57, wherein the stage is for aligning the first well receptacle or the second well receptacle with the second well receptacle.

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