Reagent degassing method

Abstract

The reagent degassing method comprises the following steps: sub-packaging a reagent into a reagent tank; performing freezing treatment on the reagent tank with the reagent so as to freeze and freeze the reagent; vacuumizing the reagent tank to form a first negative pressure in the reagent tank, and maintaining the pressure under the first negative pressure; the vacuumized reagent tank is sealed, so that the first negative pressure is maintained in the reagent tank; and transferring the sealed reagent tank into a storage environment for storage, so that the reagent is kept in a frozen state. The reagent can be prevented from generating bubbles in the use process and causing adverse effects on biochemical reaction.

Description

Technical Field

[0001] The present application relates to the field of biochemical detection, and in particular to a reagent degassing method. Background Art

[0002] During gene sequencing, the required reagents are fed into the sequencing chip through a flow system, allowing the biological samples within the chip to undergo biochemical reactions. Prior to the biochemical reaction, the reagents are typically stored in a refrigerator at approximately 4°C. The biochemical reaction process for long-chain sequencing is time-consuming, so the reagents also need to remain in the chamber for an extended period of time. The ambient temperature of the reagents is the temperature inside the chamber (often slightly above room temperature).

[0003] This means that the reagents undergo a temperature change from 4°C to room temperature before and after the biochemical reaction. With the external pressure remaining constant, the gas molecules in the reagents gradually precipitate due to the rising temperature, forming bubbles that adhere to the inner surface of the reagent tank. When bubbles accumulate at the liquid outlet of the reagent cartridge, they can be drawn into the fluidic system and potentially into the sequencing system, affecting the quality of the biochemical reaction. Summary of the Invention

[0004] In order to solve the above shortcomings, it is necessary to provide a method for degassing the reagent.

[0005] The present application provides a reagent degassing method, comprising the following steps: dispensing a reagent into a reagent tank; freezing the reagent tank containing the reagent to freeze the reagent; evacuating the reagent tank to form a first negative pressure in the reagent tank, and maintaining the pressure at the first negative pressure to make the gas content in the reagent lower than the gas saturation content of the reagent at normal temperature and pressure; sealing the vacuumed reagent tank to maintain the first negative pressure in the reagent tank; and transferring the sealed reagent tank to a storage environment for storage to keep the reagent in a frozen state.

[0006] In some possible implementations, the reagent degassing method further includes: in the storage environment, assembling the reagent tanks into a reagent kit, and packaging the reagent kit in a packaging bag to form a second negative pressure in the packaging bag.

[0007] In some possible implementations, the second negative pressure is equal to or higher than the first negative pressure.

[0008] In some possible implementations, the first negative pressure is -20 kPa to -70 kPa, and the pressure holding time is not less than 10 minutes.

[0009] In some possible implementations, the first negative pressure is -70 kPa, and the pressure holding time is 10 minutes.

[0010] In some possible implementations, the second negative pressure is -20 kPa.

[0011] In some possible implementations, the temperature of the storage environment is equal to the temperature of the freezing treatment.

[0012] In some possible implementations, vacuuming the reagent tank specifically includes: placing the frozen reagent tank into a vacuum box so that the interior of the vacuum box is connected to the interior of the reagent tank; closing the vacuum box and vacuuming the vacuum box to form the first negative pressure in the reagent tank.

[0013] In some possible implementations, the reagent tank is sealed using a plugging device. The plugging device includes a main body and an electrical control unit electrically connected to the main body, wherein the main body is placed within the vacuum chamber, and the electrical control unit is placed outside the vacuum chamber. Sealing the reagent tank specifically includes: controlling the main body to operate accordingly via the electrical control unit, thereby installing a sealing plug in the notch of the reagent tank; and depressurizing the vacuum chamber to remove the sealed reagent tank.

[0014] In some possible implementations, after the reagent tank is vacuumed, the reagent degassing method further includes: measuring the oxygen content in the reagent and converting the oxygen content into the gas content of the reagent; comparing the converted gas content with the gas saturation content of the reagent at normal temperature and pressure, so as to determine whether the gas content is lower than the gas saturation content.

[0015] The reagent degassing method of the present application is to evacuate the reagent tank after freezing the reagent so that the gas content in the reagent is lower than the gas saturation content of the reagent at normal temperature and pressure. Therefore, when the environment is under normal temperature and pressure conditions, the reagent will not separate out bubbles due to temperature increase, and thus the influence of bubbles on the flow system or sequencing system can be avoided, thereby ensuring the quality of the biochemical reaction. In addition, the present application can not only be used for deoxygenation, but also can avoid the influence of other gas components (such as nitrogen) in the reagent. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is a flow chart of a reagent degassing method provided in one embodiment of the present application.

[0017] Figure 2 for Figure 1 Schematic diagram of the structure of the reagent tank used in the method.

[0018] Figure 3 for Figure 2 Exploded view of the reagent reservoir shown.

[0019] Figure 4 for Figure 1 Module architecture diagram of the vacuum box and plugging equipment used in the method.

[0020] Figure 5 for Figure 4 The schematic diagram of the structure of the plugging device shown is after removing the electronic control part.

[0021] Figure 6 for Figure 5 The schematic diagram of the structure of the plugging device after removing the vacuum box is shown.

[0022] Figure 7 for Figure 5 The schematic diagram of the structure of the plugging device shown is from another angle after the vacuum box is removed.

[0023] Figure 8 for Figure 6 A cross-sectional view of the plugging device along line VIII-VIII.

[0024] Figure 9 for Figure 8 A cross-sectional view of the plugging device in another state.

[0025] Figure 10 for Figure 6 An exploded view of the bracket of the plugging device is shown.

[0026] Figure 11 1 is a graph showing the relationship between the oxygen content of the reagents in Example 1 and Example 2 and the test time.

[0027] Figure 12 Graph showing the relationship between the oxygen content of the reagents in Example 1 and Examples 3-4 and the test time.

[0028] Figure 13 Graph showing the relationship between the Q30 ratio in each cycle and the number of sequencing cycles in Example 1.

[0029] Figure 14 This is a graph showing the relationship between the Q30 ratio in each cycle and the number of sequencing cycles in the comparative example.

[0030] The following specific implementation methods will further illustrate the present application in conjunction with the above-mentioned drawings. DETAILED DESCRIPTION

[0031] In order to more clearly understand the above-mentioned objects, features and advantages of the present application, the present application is described in detail below in conjunction with the accompanying drawings and specific embodiments. It should be noted that, in the absence of conflict, the embodiments of the present application and the features in the embodiments can be combined with each other. In the following description, many specific details are set forth to facilitate a full understanding of the present application. The embodiments described are only a part of the embodiments of the present application, rather than all of the embodiments.

[0032] It should be noted that, although a logical order is shown in the flowchart, in some cases, the steps shown or described may be performed in an order different from that in the flowchart. The method disclosed in the embodiments of the present application includes one or more steps or actions for implementing the method. The method steps and / or actions may be interchangeable with each other without departing from the scope of the claims. Unless a specific order of steps or actions is specified, the order and / or use of specific steps and / or actions may be modified without departing from the scope of the claims.

[0033] The present application will be described below with reference to the accompanying drawings and in conjunction with embodiments.

[0034] See also Figure 1 In one embodiment of the present application, a method for degassing a reagent is provided. The order of the steps of the method may be changed, and some steps may be omitted or combined, depending on different requirements. The method comprises the following steps:

[0035] Step S1: quantitatively pack the reagent into the reagent tank A.

[0036] In some embodiments, the reagent can be prepared by mixing a powdered reagent with a solvent. The type of reagent is selected according to actual needs, for example, the reagent can be a thermal reaction reagent (HOT reagent) or a removal reaction reagent (CMR reagent).

[0037] like Figure 2 and Figure 3 As shown, in some embodiments, a reagent tank A comprises a tank body A1, which is provided with at least one notch A2 for communication with the outside world. Tank body A1 may include a tank body A10 and an encapsulating film A11 encapsulating the sidewalls of tank body A10. Reagent tank A is a dual-port reagent tank, with a notch A2 defined on opposite sides of tank body A10. During dispensing, reagents can be injected into reagent tank A through one of the notches A2.

[0038] Step S2: freezing the reagent tank A containing the reagent, so that the reagent in the reagent tank A is frozen.

[0039] In some embodiments, the reagent tank A containing the reagent can be placed in a predetermined temperature and frozen for a period of time. The predetermined temperature is -20°C, and the freezing time is not less than 12 hours.

[0040] In step S3, the reagent tank A after freezing is vacuumed to form a first negative pressure in the reagent tank A, and then the first negative pressure is maintained for a period of time.

[0041] In the present application, by vacuumizing the reagent tank A after freezing and maintaining pressure for a period of time, the gas molecules in the reagent are separated from the reagent, thereby reducing the gas content in the reagent. After the above-mentioned vacuum treatment, the gas content in the reagent is lower than the gas saturation content of the reagent at normal temperature and pressure. It will be understood that the gas saturation content of the reagent at normal temperature and pressure refers to that when the environment is under normal temperature and pressure, the gas content dissolved in the reagent from the environment and the gas content deviated from the reagent to the environment reach a dynamic equilibrium. Therefore, when the environment is under normal temperature and pressure, the reagent will not separate out bubbles because of the temperature increase, and therefore the influence of bubbles on the flow system or sequencing system can be avoided, thereby ensuring the quality of the biochemical reaction.

[0042] A pH meter can be used to measure the oxygen content in the reagent. Since air contains approximately 20% oxygen molecules, the oxygen content measured by the pH meter can be converted to the gas content in the reagent. The converted gas content in the reagent is then compared with the saturated gas content of the reagent at room temperature and pressure to determine whether the gas content in the reagent is lower than the saturated gas content of the reagent at room temperature and pressure.

[0043] In some embodiments, the first negative pressure is between -20 kPa and -70 kPa, and the holding time is no less than 10 minutes. The lower the first negative pressure, the better the degassing effect, and the required holding time can be appropriately shortened. In some specific embodiments, the first negative pressure is -70 kPa, and the holding time is 10 minutes. This negative pressure condition not only reduces the gas content in the reagent to below the gas saturation content of the reagent at room temperature and pressure, but also helps shorten production time and improve production efficiency.

[0044] In some embodiments, the frozen reagent tank A is first placed in a vacuum box, which is then closed and evacuated. In this step, the interior of the vacuum box is required to be connected to the interior of the reagent tank A. Due to the evacuation of the vacuum box, a negative pressure is generated inside the reagent tank A.

[0045] In step S4, the vacuumed reagent tank A is sealed so that the first negative pressure is maintained in the reagent tank A.

[0046] The reagent is sealed in the reagent tank A. At this time, the negative pressure in the reagent tank A is maintained and the reagent is basically not thawed.

[0047] In some embodiments, a sealing plug is installed in the notch A2 of the reagent tank A by a pressing device, thereby sealing the reagent tank A. Figure 4Module diagram of the tampon equipment and vacuum box in some embodiments of the present application. The tampon equipment 100 includes a main body 1 and an electric control part 2 electrically connected to the main body 1. The main body 1 of the tampon equipment 100 is placed in the vacuum box 200 of step S3, and the electric control part 2 is placed outside the vacuum box 200. During specific operation, after the vacuum box 200 is vacuumed, the main body 1 is controlled by the electric control part 2 of the tampon equipment 100 to close the notch of the reagent tank A, then the vacuum box 200 is depressurized, and then the reagent tank A after sealing is taken out. The inside of the reagent tank A after sealing can now maintain a negative pressure state. In certain embodiments, the vacuum box 200 and the tampon equipment 100 can be independent of each other, and the tampon equipment 100 can be placed in the vacuum box 200 during use. In other embodiments, the vacuum box 200 and the tampon equipment 100 can also be made into a whole.

[0048] Please refer to Figures 5 to 8 The main body 1 of the plugging device 100 includes a bracket 10, a movable plate 20, a pressing member 30, and a driving member 40. The electronic control unit 2 of the plugging device 100 is electrically connected to the driving member 40 to control the operation of the driving member 40. The electronic control unit 2 can be a circuit board, a chip, or other type of controller.

[0049] The bracket 10 includes a first frame 11 and a second frame 12 stacked together, with the first frame 11 located above the second frame 12. The first frame 11 and the second frame 12 are detachably mounted together. Figure 10 The first frame 11 has at least one first slot 110, each of which is configured to receive a sealing plug B. The second frame 12 has at least one second slot 120, each of which is configured to receive a chilled reagent reservoir A. The first slot 110 and the second slot 120 are arranged in a corresponding direction D1 between the first frame 11 and the second frame 12. In some embodiments, the sealing plug B is made of an elastic material (e.g., rubber). In a direction D2 perpendicular to direction D1, the width of the first slot 110 is smaller than the width of the sealing plug B, allowing the sealing plug B to be installed in the first slot 110 via an interference fit. In some embodiments, there are at least two first slots 110 and two second slots 120, each of which is configured to receive a corresponding direction D1. In some embodiments, the bracket 10 may further include a base plate 13 and a support frame 14 secured to the base plate 13. The second frame 12 is secured to the base plate 13 via the support frame 14.

[0050] During operation, the first frame 11 and the second frame 12 can be separated, and then the sealing plug B and the reagent tank A can be installed in the first slot 110 and the second slot 120 respectively. Then the first frame 11 can be stacked and installed on top of the second frame 12. At this time, the sealing plug B is located above the notch A2 of the reagent tank A. Figure 8 As shown, a gap G is provided between the sealing plug B and the notch A2 of the reagent tank A. More specifically, a gap G is provided between the outer peripheral surface of the bottom of the sealing plug B and the inner wall of the notch A2. This gap G is used to connect the interior spaces of the vacuum box 200 and the reagent tank A, so that the same negative pressure is generated inside the reagent tank A after the vacuum box 200 is evacuated. In some embodiments, the width W of the gap G in the direction D2 is not less than 1 mm.

[0051] The movable plate 20 is disposed above the bracket 10 and is connected to the driving member 40. The pressing member 30 is fixed to the surface of the movable plate 20 facing the bracket 10 and protrudes from the surface of the movable plate 20 facing the bracket 10. In the direction D1, the pressing member 30 is disposed corresponding to the first slot 110. Figure 8 As shown, when the driving member 40 does not drive the moving plate 20 to move, the main body 1 is in the first state, at which time a gap G is provided between the outer peripheral surface of the bottom of the sealing plug B and the inner wall of the notch A2. Figure 9 As shown, after the vacuum box 200 is evacuated, the driving member 40 is used to drive the movable plate 20 toward the bracket 10, thereby driving the pressing member 30 to push the sealing plug B in the first slot 110, so that the sealing plug B is pushed into the notch A2 of the reagent tank A in the second slot 120, thereby achieving a seal. At this time, the main body 1 is in the second state. Thereafter, the driving member 40 is used to drive the movable plate 20 to move back to the initial position, returning the main body 1 to the first state. In some embodiments, the driving member 40 is a motor.

[0052] In some embodiments, the main body 1 of the plugging device 100 may further include a first fixing plate 50 and a second fixing plate 60, and the movable plate 20 and the bracket 10 are both arranged between the first fixing plate 50 and the second fixing plate 60. The bracket 10 is mounted on the first fixing plate 50. The second fixing plate 60 is stacked above the movable plate 20, that is, the movable plate 20 is arranged between the second fixing plate 60 and the bracket 10. The second fixing plate 60 is provided with a through hole 61 (at Figure 8 The driving member 40 is mounted on the second fixed plate 60. Specifically, the driving member 40 is located on the surface of the second fixed plate 60 facing away from the movable plate 20. The driving end of the driving member 40 passes through the through hole 61 and is connected to the movable plate 20, thereby driving the movable plate 20 to move.

[0053] Further, if Figure 7As shown, in some embodiments, the main body 1 of the plugging device 100 may further include a fixed seat 80, a first optical coupling assembly 81, and a second optical coupling assembly 82. The fixed seat 80 is fixed to the first fixed plate 50, and the first optical coupling assembly 81 and the second optical coupling assembly 82 are both fixed to the fixed seat 80. Moreover, the first optical coupling assembly 81 is located above the second optical coupling assembly 82 in the direction D1. Each of the first optical coupling assembly 81 and the second optical coupling assembly 82 includes a first end 811 and a second end 812 arranged opposite to each other in the direction D2, and the first end 811 is used to emit light toward the second end 812. A baffle 21 is also provided on the movable plate 20. The baffle 21 is selectively inserted between the first end 811 and the second end 812 of the first optical coupling assembly 81 or between the first end 811 and the second end 812 of the second optical coupling assembly 82 when the movable plate 20 moves, thereby blocking the light emitted by the first end 811. More specifically, when the main body 1 is in the first state, the baffle 21 is inserted between the first end 811 and the second end 812 of the first optical coupling assembly 81. At this time, the second end 812, not receiving the light emitted by the first end 811, sends a corresponding feedback signal to the electronic control unit 2. Similarly, when the main body 1 is in the second state, the baffle 21 is inserted between the first end 811 and the second end 812 of the second optical coupling assembly 82. At this time, the second end 812, not receiving the light emitted by the first end 811, also sends a corresponding feedback signal to the electronic control unit 2. Thus, the electronic control unit 2 can determine whether the main body 1 is in the first state or the second state based on whether the feedback signal is received, thereby controlling the driver 40 to stop operating. In other embodiments, the electronic control unit 2 can also control the driver 40 to move a predetermined number of steps, thereby moving the movable plate 20 and the pressing member 30 fixed to the movable plate 20 a corresponding distance. After the pressing member 30 has moved the corresponding distance, the main body 1 switches to the second state or the first state.

[0054] Furthermore, in some embodiments, at least one pad 51 may be provided on the surface of the first fixing plate 50 facing away from the second fixing plate 60. The pad 51 may be made of elastic material (such as rubber) to reduce wear of the main body 1 of the plugging device 100. Figure 6 and Figure 7 As shown, in some embodiments, a guide groove 52 is provided on the surface of the first fixing plate 50 facing the second fixing plate 60. A guide block 111 is provided on the surface of the bracket 10 facing the first fixing plate 50 (e.g., a guide block 111 is provided on the bottom plate 13). The bracket 10 can be slidably mounted on the first fixing plate 50 through the cooperation between the guide block 111 and the guide groove 52, thereby facilitating the mounting of the bracket 10 on the first fixing plate 50. A positioning block 53 may also be provided on the surface of the first fixing plate 50 facing the second fixing plate 60. The positioning block 53 is provided at one end of the guide groove 52 and is used to position the bracket 10 after the bracket 10 slides to a certain position.

[0055] Furthermore, in some embodiments, the main body 1 of the plugging device 100 may also include at least one guide rod 70. Each guide rod 70 is connected between the first fixed plate 50 and the second fixed plate 60 and extends along the direction D1. The guide rod 70 is also slidably disposed through the movable plate 20, so that when the movable plate 20 moves toward or away from the bracket 10, the guide rod 70 can serve as a guide and prevent the movable plate 20 from deviating from its direction of movement. In some specific embodiments, there are four guide rods 70, each of which passes through the four corners of the movable plate 20.

[0056] Step S5: Transfer the sealed reagent tank A to a storage environment to keep the reagent in the reagent tank A in a frozen state.

[0057] In some embodiments, the reagent tank A can be transferred to a storage environment having the predetermined temperature to maintain the reagent in the reagent tank A in a frozen state. In some specific embodiments, the storage environment is a cold storage with a temperature of -20°C.

[0058] In step S6, the reagent tank A is assembled into a reagent kit in this storage environment, and the reagent kit is then packaged in a packaging bag using a vacuum packaging machine to form a second negative pressure in the packaging bag. The second negative pressure can be equal to or higher than the first negative pressure.

[0059] The vacuum packaging machine is used to extract air from the packaging bag and complete the sealing process after achieving a predetermined vacuum level. Because the test kit is vacuum-packed, it provides negative pressure protection. Even if there is a leak in reagent reservoir A, a certain negative pressure condition can be maintained. Furthermore, the storage environment of the test kit is maintained at the predetermined temperature, so the reagents in reagent reservoir A remain frozen.

[0060] In some embodiments, when the first negative pressure is -70 kPa, the second negative pressure is -20 kPa.

[0061] Step S7: Place the packaging bag containing the test kit into a paper packaging box.

[0062] Later, when the reagents need to be used, the test kit is first transferred to a refrigerator at 4°C to thaw the reagents in the frozen state in the test kit, and then the reagents are introduced into the sequencer for reaction. The internal temperature of the sequencer is room temperature. During this process, the reagents will successively pass through three temperatures: -20°C (storage temperature), 4°C (thawing temperature), and 25°C (working temperature). When the reagent is thawed at a temperature of 4°C, the air content in the reagent is lower than the gas saturation content of the reagent at the working temperature, thereby avoiding the phenomenon of gas precipitation and formation of bubbles in the reagent due to the increase in ambient temperature, thereby avoiding the above-mentioned bubbles affecting the quality of the biochemical reaction. It can be understood that the above-mentioned storage temperature, thawing temperature and working temperature are for illustration only, and they can all be changed according to actual conditions.

[0063] Moreover, since the reagents of the present application are in a negative pressure state from the production end (referring to the step of evacuating the frozen reagent tank A in step S3) to the storage end (referring to the step of placing the packaging bag with the reagent kit into the packaging box in step S7), it can be fully guaranteed that the reagents will not be separated into bubbles due to the increase in temperature during subsequent use. In addition, compared with the existing freeze-thaw method that introduces an inert gas (such as nitrogen) to remove the oxygen contained in the reagent, the present application can not only be used for deoxygenation, but also avoid the influence of other gas components (such as nitrogen) in the reagent.

[0064] The reagent degassing method provided in this application is described below by using specific examples and comparative examples. Those skilled in the art should understand that the reagent degassing method described in this application is only an example, and any other suitable reagent degassing method is within the scope of this application.

[0065] Example 1

[0066] Two reagent tanks containing 22 mL of HOT reagent and 22 mL of CMR reagent, respectively, were placed in a freezer at -20°C for 24 hours to freeze the reagents in the reagent tanks. The frozen reagent tanks were placed in a vacuum box (manufacturer: Tongrun; model: VB-1) and evacuated using a vacuum pump (manufacturer: Tingwei; model: TW-4A) to a pressure of -70 kPa in the reagent tanks. The pressure was then maintained at this pressure for 10 minutes. The evacuated reagent tanks were sealed and transferred to a cold storage at -20°C. The reagent tanks were then assembled into a test kit. The test kits were then packaged in a packaging bag using a vacuum packaging machine (manufacturer: Chuangmeng; model: VS600) to a pressure of -20 kPa in the packaging bag. The test kits were then frozen and stored in the cold storage for 24 hours. The reagent tanks were then thawed in a refrigerator at 4°C for 4 hours.

[0067] Example 2

[0068] The difference from Example 1 is that the reagent tank is evacuated to a negative pressure of -70 kPa, and then maintained at this negative pressure for 2 hours and 30 minutes.

[0069] The reagent tanks of Example 1 and Example 2 were respectively turned upside down 10 times for mixing (simulating the mixing treatment of the reagents before the actual machine reaction), and allowed to stand for 10 minutes. Then, under normal temperature and pressure conditions, the temperature change and oxygen content change of the reagents in the reagent tank were measured as the test time was extended (simulating the reaction time during the actual machine reaction). The oxygen content can be measured by a pH meter, and the measurement results are recorded in Table 1.

[0070] Table 1

[0071]

[0072] Then, the saturated oxygen content of each reagent was tested at different ambient temperatures under normal pressure. It is understood that after a reagent is left at a certain ambient temperature for a period of time, the amount of gas dissolved in the reagent from the environment and the amount of gas released from the reagent into the environment reach a dynamic equilibrium. The oxygen content of the reagent measured at this time is the saturated oxygen content of the reagent. 24°C is room temperature, and 30°C and 35°C are both reagent temperatures achieved in a water bath. The measurement results are recorded in Table 2.

[0073] Table 2

[0074]

[0075] The relationship between the oxygen content of the reagents in Example 1 and Example 2 and the test time is plotted according to Table 1. The results are shown in Figure 11 Further, the saturated oxygen content of the reagents at 24°C and 30°C measured in Table 2 was added to Figure 11 Among them, 24℃ is the normal working temperature of the reagent during sequencing, and 30℃ can be regarded as the maximum working temperature of the reagent during sequencing. Figure 11 It can be seen that the oxygen content of the reagent in Example 1 is higher than that in Example 2, indicating that the oxygen content in the reagent decreases with the extension of the pressure holding time. At the same time, the oxygen content of the reagent in Example 1 and Example 2 is less than its saturated oxygen content within 20 hours, indicating that a pressure holding time of 10 minutes can meet actual needs.

[0076] Example 3

[0077] The difference from Example 1 is that after the vacuum box is evacuated by a vacuum pump, the negative pressure in the reagent tank is -30 kPa.

[0078] Example 4

[0079] The difference from Example 1 is that after the vacuum box is evacuated by a vacuum pump, the negative pressure in the reagent tank is -20 kPa.

[0080] Similar steps were used to measure the temperature change and oxygen content change of the reagents of Examples 3-4 as the test time increased under normal temperature and pressure conditions. The measurement results are recorded in Table 3.

[0081] Table 3

[0082]

[0083] The relationship between the oxygen content of the reagents in Example 1 and Example 3-4 and the test time is plotted according to Table 3. The results are shown in Figure 12Further, the saturated oxygen content of the reagents at 24°C and 30°C measured in Table 2 was added to Figure 12 Among them, 24℃ is the normal working temperature of the reagent during sequencing, and 30℃ can be regarded as the maximum working temperature of the reagent during sequencing. Figure 12 It can be seen that the oxygen content of the reagents in Examples 3-4 is less than its saturated oxygen content within 20 hours. At the same time, the oxygen content of the reagents in Examples 3-4 is higher than that of the reagent in Example 1, indicating that the negative pressure condition of -70 kPa is more conducive to ensuring the degassing quality of the reagents.

[0084] Comparative Example

[0085] The difference from Example 1 is that the reagent tank is not vacuumed after being frozen, that is, the frozen reagent tank is directly sealed and transferred to a cold storage at a temperature of -20°C for frozen storage for 24 hours.

[0086] The reagent tanks of Example 1 and the comparative example were subjected to PE150 sequencing on a sequencing platform (manufacturer: BGI; model: DNBSEQ-E25 machine), and the chip model was DNBSEQ-E25 chip. The Q30 (%) decrease in each cycle was then counted to evaluate the sequencing quality. Q30 is a quality value for sequencing. Mathematically speaking, Q30 means that the probability of misidentification is 0.1%, that is, the error rate is 0.1%, or the accuracy is 99.9%. In sequencing evaluation, in order to evaluate the accuracy of sequencing reads, Q30 and the proportion of all base quality values ​​greater than Q30 will be evaluated. The higher the Q30 value, the better the sequencing quality. Among them, from Figure 13 and Figure 14 It can be seen that compared with Example 1, the comparative example caused a sudden drop of about 3% in Q30 after a certain cycle due to bubbles entering the sequencing system and could not be recovered later, indicating that the reagents in the comparative example affected the sequencing quality due to the precipitation of bubbles.

[0087] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present application and are not intended to limit the present application. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present application.

Claims

1. A reagent degassing method, characterized in that: The steps include: Dispense the reagents into the reagent tank; Freezing the reagent tank containing the reagent to freeze the reagent; The reagent tank is subjected to a vacuum treatment to form a first negative pressure in the reagent tank, and the pressure is maintained at the first negative pressure so that the gas content in the reagent is lower than the gas saturation content of the reagent at normal temperature and pressure; Sealing the vacuumed reagent tank to maintain the first negative pressure in the reagent tank; The sealed reagent tank is transferred to a storage environment for storage so that the reagent remains in a frozen state.

2. The reagent degassing method according to claim 1, characterized in that: Also includes: In the storage environment, the reagent tanks are assembled into a reagent kit, and the reagent kit is packaged in a packaging bag, so that a second negative pressure is formed in the packaging bag.

3. The reagent degassing method according to claim 2, characterized in that: The second negative pressure is equal to or higher than the first negative pressure.

4. The reagent degassing method according to claim 3, characterized in that: The first negative pressure is -20 kPa to -70 kPa, and the pressure maintaining time is not less than 10 minutes.

5. The reagent degassing method according to claim 4, characterized in that: The first negative pressure is -70 kPa, and the pressure-maintaining time is 10 minutes.

6. The reagent degassing method according to claim 5, characterized in that: The second negative pressure is -20 kPa.

7. The reagent degassing method according to claim 1, characterized in that: The temperature of the storage environment is equal to the temperature of the freezing process.

8. The reagent degassing method according to claim 1, characterized in that: The vacuum treatment of the reagent tank specifically includes: placing the frozen reagent tank into a vacuum box so that the interior of the vacuum box is connected to the interior of the reagent tank; The vacuum box is closed and evacuated to form the first negative pressure in the reagent tank.

9. The reagent degassing method according to claim 8, characterized in that: The reagent tank is sealed in a plugging device, the plugging device includes a main body and an electric control part electrically connected to the main body, the main body is placed in the vacuum box, and the electric control part is placed outside the vacuum box. The reagent tank is sealed specifically including: The electrical control part controls the main body part to operate accordingly, thereby installing the sealing plug in the notch of the reagent tank; The vacuum box is depressurized and the sealed reagent tank is taken out.

10. The reagent degassing method according to claim 1, characterized in that: After the reagent tank is vacuumed, the reagent degassing method further comprises: measuring the oxygen content in the reagent and converting the oxygen content into the gas content of the reagent; The converted gas content is compared with the gas saturation content of the reagent at normal temperature and pressure to determine whether the gas content is lower than the gas saturation content.

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