Reagent degassing method

Abstract

The application discloses a reagent degassing method, which comprises the following steps: reagent is divided into reagent tanks; the reagent tanks with the reagents are subjected to freezing treatment to freeze the reagents into ice; the reagent tanks are subjected to vacuumizing treatment to form a first negative pressure in the reagent tanks and keep the first negative pressure; the reagent tanks after the vacuumizing treatment are sealed to maintain the first negative pressure in the reagent tanks; and the sealed reagent tanks are transferred to a storage environment for storage, so that the reagents are maintained in a frozen state. The application can avoid the generation of bubbles in the reagents during use and the adverse effects on biochemical reactions.

Description

Technical Field

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

[0002] During gene sequencing, the necessary reagents are fed into the sequencing chip via a flow path system, causing the biological sample within the chip to undergo corresponding biochemical reactions. Before the biochemical reactions, the reagents are generally stored in a refrigerator at around 4°C. The biochemical reaction process for long-chain sequencing is time-consuming, therefore the reagents also need to be placed in the chamber for an extended period during the biochemical reactions, and the ambient temperature of the reagents is the temperature inside the chamber (often slightly higher than room temperature).

[0003] In other words, the reagents undergo a temperature change from 4°C to room temperature before and after the biochemical reaction. Under constant external pressure, gas molecules in the reagents will gradually precipitate out as the temperature rises, adhering to the inner surface of the reagent tank in the form of bubbles. When bubbles accumulate at the reagent kit outlet, they may be drawn into the flow path system and potentially into the sequencing system, affecting the quality of the biochemical reaction. Summary of the Invention

[0004] To address the above shortcomings, it is necessary to provide a method for degassing reagents.

[0005] This application provides a reagent degassing method, comprising the following steps: dispensing the reagent into a reagent tank; freezing the reagent tank containing the reagent to freeze the reagent; evacuating the reagent tank to create a first negative pressure, and maintaining the pressure under the first negative pressure to ensure that the gas content in the reagent is lower than the gas saturation content of the reagent at room temperature and pressure; sealing the evacuated reagent tank to maintain the first negative pressure; 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: assembling the reagent tank into a reagent kit in the storage environment, and encapsulating the reagent kit in a packaging bag to create a second negative pressure inside 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 between -20 kPa and -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 process.

[0012] In some possible implementations, evacuating the reagent tank specifically includes: placing the frozen reagent tank into a vacuum chamber, making the interior of the vacuum chamber connected to the interior of the reagent tank; closing the vacuum chamber and evacuating it to create the first negative pressure inside the reagent tank.

[0013] In some possible implementations, the reagent tank is sealed using a stoppering device. The stoppering device includes a main body and an electrical control unit electrically connected to the main body. The main body is placed inside the vacuum chamber, and the electrical control unit is located outside the vacuum chamber. Sealing the reagent tank specifically includes: controlling the main body to operate via the electrical control unit, thereby installing a sealing plug into the opening of the reagent tank; depressurizing the vacuum chamber, and removing the sealed reagent tank.

[0014] In some possible implementations, after evacuating the reagent bath, 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 room temperature and pressure to determine whether the gas content is lower than the gas saturation content.

[0015] The reagent degassing method of this application involves freezing the reagent and then evacuating the reagent bath to reduce the gas content in the reagent below its saturation level at room temperature and pressure. Therefore, when the environment is at room temperature and pressure, the reagent will not precipitate bubbles due to temperature rise, thus avoiding the impact of bubbles on the flow path system or sequencing system, thereby ensuring the quality of the biochemical reaction. Furthermore, this application can not only be used for deoxygenation but also avoids the influence of other gaseous components in the reagent (such as nitrogen). Attached Figure Description

[0016] Figure 1 A flowchart of a reagent degassing method provided in one embodiment of this application.

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

[0018] Figure 3 for Figure 2 An exploded view of the reagent tank shown.

[0019] Figure 4 for Figure 1 The method uses a modular architecture diagram of the vacuum chamber and plugging equipment.

[0020] Figure 5 for Figure 4 The diagram shown is a structural schematic of the plugging device after the electrical control components have been removed.

[0021] Figure 6 for Figure 5 The diagram shows the structure of the plugging device after removing the vacuum chamber.

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

[0023] Figure 8 for Figure 6 A cross-sectional view of the plugging device along 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 The exploded view of the support frame of the plugging device is shown.

[0026] Figure 11 The graph shows the relationship between the oxygen content of the reagents and the test time in Examples 1 and 2.

[0027] Figure 12 The graph shows the relationship between the oxygen content of the reagents and the test time in Examples 1 and 3-4.

[0028] Figure 13 This is a graph showing the relationship between the proportion of Q30 in each cycle and the number of sequencing cycles in Example 1.

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

[0030] The following detailed description, in conjunction with the accompanying drawings, will further illustrate this application. Detailed Implementation

[0031] To better understand the above-mentioned objectives, features, and advantages of this application, the application will be described in detail below with reference to the accompanying drawings and specific embodiments. It should be noted that, unless otherwise specified, the embodiments and features described in the embodiments of this application can be combined with each other. Many specific details are set forth in the following description to provide a thorough understanding of this application; the described embodiments are only some embodiments of this application, and not all 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 a different order than that shown in the flowchart. The methods disclosed in the embodiments of this application include one or more steps or actions for implementing the method. Method steps and / or actions may be interchanged 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 now be described with reference to the accompanying drawings and embodiments.

[0034] Please see Figure 1 This application provides a reagent degassing method according to one embodiment. Depending on different needs, the order of steps in the above method can be changed, and some steps can be omitted or combined. The method includes the following steps:

[0035] Step S1: Dispense the reagent quantitatively into 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 above-mentioned reagent can be a thermal reaction reagent (HOT reagent) or a cleavage reaction reagent (CMR reagent).

[0037] like Figure 2 and Figure 3 As shown, in some embodiments, reagent tank A includes a tank body A1, with at least one opening A2 communicating with the outside. The tank body A1 may include a tank body A10 and an encapsulating film A11 sealed to the sidewall of the tank body A10. The reagent tank A is a double-port reagent tank, with an opening A2 on each of the opposite sides of the tank body A10. During dispensing, reagent can be injected into the reagent tank A through one of the openings A2.

[0038] Step S2: Freeze the reagent tank A containing the reagent to freeze the reagent in the reagent tank A.

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

[0040] Step S3: Vacuum the frozen reagent tank A to create a first negative pressure inside the reagent tank A, and then maintain the pressure under the first negative pressure for a period of time.

[0041] In this application, by evacuating and pressurizing the frozen reagent tank A for a period of time, gas molecules in the reagent are separated from the reagent, thereby reducing the gas content in the reagent. After the above vacuum treatment, the gas content in the reagent is lower than the gas saturation content of the reagent at room temperature and pressure. It can be understood that the gas saturation content of the reagent at room temperature and pressure refers to the dynamic equilibrium between the amount of gas dissolved in the reagent from the environment and the amount of gas released from the reagent into the environment under normal temperature and pressure conditions. Therefore, when the environment is at normal temperature and pressure, the reagent will not precipitate bubbles due to temperature increase, thus avoiding the impact of bubbles on the flow path system or sequencing system, thereby ensuring the quality of the biochemical reaction.

[0042] The oxygen content in the reagent can be measured using a pH meter. Since air contains approximately 20% oxygen molecules, the oxygen content measured by the pH meter can be converted into the gas content of the reagent. Then, the converted gas content is compared with the reagent's gas saturation content at room temperature and pressure to determine whether the gas content is lower than the reagent's gas saturation content 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 not 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 ensures that the gas content in the reagent is lower than the gas saturation content of the reagent at room temperature and pressure, but also helps to shorten the production time and improve production efficiency.

[0044] In some embodiments, the frozen reagent tank A is first placed in a vacuum chamber, the vacuum chamber is closed, and a vacuum is evacuated. During this step, the interior of the vacuum chamber must be in communication with the interior of the reagent tank A. Due to the vacuum evacuation process, a negative pressure is created inside the reagent tank A.

[0045] Step S4: Seal the reagent tank A after vacuuming to maintain the first negative pressure inside the reagent tank A.

[0046] In this process, the reagent is sealed in reagent tank A, where the negative pressure inside reagent tank A is maintained and the reagent is basically not thawed.

[0047] In some embodiments, a sealing plug is installed in the slot A2 of reagent tank A by a plugging device, thereby sealing reagent tank A. Figure 4This is a block diagram of the stoppering device and vacuum chamber in some embodiments of this application. The stoppering device 100 includes a main body 1 and an electrical control part 2 electrically connected to the main body 1. The main body 1 of the stoppering device 100 is placed inside the vacuum chamber 200 in step S3, while the electrical control part 2 is placed outside the vacuum chamber 200. In specific operation, after the vacuum chamber 200 is evacuated, the electrical control part 2 of the stoppering device 100 controls the main body 1 to seal the opening of the reagent tank A, then the vacuum chamber 200 is depressurized, and then the sealed reagent tank A is removed. At this time, the inside of the sealed reagent tank A can maintain a negative pressure state. In some embodiments, the vacuum chamber 200 and the stoppering device 100 can be independent of each other, and the stoppering device 100 can be placed inside the vacuum chamber 200 during use. In other embodiments, the vacuum chamber 200 and the stoppering device 100 can also be made into a single unit.

[0048] Please refer to the above as well. Figures 5 to 8 The main body 1 of the compression device 100 includes a support 10, a movable plate 20, a pressing component 30, and a driving component 40. The electrical control unit 2 of the compression device 100 is electrically connected to the driving component 40 to control its corresponding operations. The electrical control unit 2 can be a circuit board, a chip, or other type of controller.

[0049] The support frame 10 includes a first frame 11 and a second frame 12 stacked together, with the first frame 11 positioned above the second frame 12. The first frame 11 and the second frame 12 are detachably mounted together. Please refer to [reference needed]. Figure 10 The first frame 11 has at least one first slot 110, each first slot 110 being used to install a sealing plug B. The second frame 12 has at least one second slot 120, each second slot 120 being used to install a frozen reagent tank A. The first slots 110 and the second slots 120 are correspondingly arranged in the stacking direction D1 of the first frame 11 and the second frame 12. In some embodiments, the sealing plug B is made of an elastic material (such as rubber), and in a direction D2 perpendicular to the D1 direction, the width of the first slot 110 is smaller than the width of the sealing plug B, so that the sealing plug B can be installed in the first slot 110 by an interference fit. In some embodiments, the number of first slots 110 and second slots 120 is at least two, and the two first slots 110 and the two second slots 120 are correspondingly arranged in the D1 direction. In some embodiments, the bracket 10 may further include a base plate 13 and a support frame 14 fixed to the base plate 13, and the second frame 12 is fixed to the base plate 13 by the support frame 14.

[0050] During operation, the first frame 11 and the second frame 12 can be separated. Then, the sealing plug B and the reagent tank A are installed in the first slot 110 and the second slot 120 respectively. The first frame 11 is then stacked and installed on top of the second frame 12, with the sealing plug B positioned above the slot opening A2 of the reagent tank A. At this point, as... Figure 8 As shown, a gap G is provided between the sealing plug B and the opening 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 opening A2. This gap G is used to connect the internal spaces of the vacuum chamber 200 and the reagent tank A, so that the same negative pressure is generated inside the reagent tank A after the vacuum chamber 200 is evacuated. In some embodiments, the width W of the gap G in the D2 direction is not less than 1 mm.

[0051] The movable plate 20 is positioned above the bracket 10 and connected to the drive member 40. The pressing member 30 is fixed to the surface of the movable plate 20 facing the bracket 10 and protrudes from this surface. In the D1 direction, the pressing member 30 corresponds to the first groove 110. Figure 8 As shown, when the driving component 40 does not drive the moving plate 20 to move, the main body 1 is in the first state, and at this time, there is a gap G between the outer peripheral surface of the bottom of the sealing plug B and the inner wall of the groove A2. Figure 9 As shown, after the vacuum chamber 200 is evacuated, the drive unit 40 drives the moving plate 20 to move towards the support 10, thereby causing 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 slot 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. Afterwards, the drive unit 40 drives the moving plate 20 to move back to the initial position, so that the main body 1 returns to the first state. In some embodiments, the drive unit 40 is a motor.

[0052] In some embodiments, the main body 1 of the compression device 100 may further include a first fixing plate 50 and a second fixing plate 60, with a movable plate 20 and a bracket 10 both disposed 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 on top of the movable plate 20, that is, the movable plate 20 is disposed between the second fixing plate 60 and the bracket 10. The second fixing plate 60 is provided with a through hole 61 (in... Figure 8 (As shown in the diagram). The drive member 40 is mounted on the second fixed plate 60, specifically located on the surface of the second fixed plate 60 opposite to the movable plate 20. The drive end of the drive 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] Furthermore, such as Figure 7As shown, in some embodiments, the main body 1 of the compression device 100 may further include a fixed base 80, a first optical coupler assembly 81, and a second optical coupler assembly 82. The fixed base 80 is fixed to the first fixed plate 50, and both the first optical coupler assembly 81 and the second optical coupler assembly 82 are fixed to the fixed base 80. Moreover, the first optical coupler assembly 81 is located above the second optical coupler assembly 82 in the D1 direction. Each of the first optical coupler assembly 81 and the second optical coupler assembly 82 includes a first end 811 and a second end 812 disposed opposite to each other in the D2 direction, the first end 811 being used to emit light toward the second end 812. The movable plate 20 is also provided with a baffle 21, which selectively inserts between the first end 811 and the second end 812 of the first optical coupler assembly 81 or between the first end 811 and the second end 812 of the second optical coupler 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 optocoupler assembly 81. Since the second end 812 does not receive light emitted from the first end 811, it 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 optocoupler assembly 82. Since the second end 812 does not receive light emitted from the first end 811, it 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 or second state based on whether a feedback signal is received, thereby controlling the drive unit 40 to stop operating. In other embodiments, the electronic control unit 2 can also control the drive unit 40 to move a predetermined number of steps, thereby moving the moving plate 20 and the pressing member 30 fixed to the moving plate 20 a corresponding distance. After the pressing member 30 has moved the corresponding distance, the main body 1 switches to the second or first state.

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

[0055] Furthermore, in some embodiments, the main body 1 of the compression 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 D1 direction. The guide rod 70 also slidably passes through the movable plate 20, so that when the movable plate 20 moves toward or away from the support 10, the guide rod 70 can guide it and prevent the moving direction of the movable plate 20 from deviating. In some specific embodiments, there are four guide rods 70, which pass through the four corners of the movable plate 20 respectively.

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

[0057] In some embodiments, reagent tank A can be transferred to a storage environment with the predetermined temperature described above, so that the reagents in reagent tank A remain frozen. In some specific embodiments, the storage environment is a cold storage room with a temperature of -20°C.

[0058] Step S6: In this storage environment, reagent tank A is assembled into a reagent kit, and then the reagent kit is sealed in a packaging bag using a vacuum packaging machine, creating a second negative pressure inside 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 remove air from the packaging bag, achieving a predetermined vacuum level before sealing. Because the reagent kit is vacuum-packed, negative pressure is maintained, ensuring a certain level of negative pressure even if there is a leak in reagent tank A. Furthermore, the reagent kit is stored at the predetermined temperature, thus keeping the reagents in reagent tank A 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 reagent kit into the paper packaging box.

[0062] When reagents are needed subsequently, the kit is first transferred to a 4°C freezer to thaw the frozen reagents. Then, the reagents are introduced into the sequencer for reaction, which operates at room temperature. During this process, the reagents are successively subjected to three temperatures: -20°C (storage temperature), 4°C (thawing temperature), and 25°C (operating temperature). When the reagents thaw at 4°C, the air content is lower than the gas saturation level at the operating temperature. This prevents gas from escaping and forming bubbles due to increased ambient temperature, thus avoiding any impact on the quality of the biochemical reaction. It should be understood that the storage temperature, thawing temperature, and operating temperature mentioned above are illustrative and can be adjusted according to actual conditions.

[0063] Furthermore, since the reagents in this application are kept under negative pressure throughout the entire process from production (referring to the step of evacuating the frozen reagent tank A in step S3) to storage (referring to the step of placing the packaging bag containing the reagent kit into the packaging box in step S7), it can be fully guaranteed that the reagents will not release gas and form bubbles due to temperature rise during subsequent use. In addition, compared with the existing freeze-thaw method of introducing an inert gas (such as nitrogen) to remove the oxygen contained in the reagents, this application can not only be used for deoxygenation, but also avoid the influence of other gaseous components (such as nitrogen) in the reagents.

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

[0065] Example 1

[0066] Two reagent trays, each containing 22 mL of HOT reagent and 22 mL of CMR reagent respectively, were frozen at -20°C for 24 hours to allow the reagents to solidify. The frozen reagent trays were then placed in a vacuum chamber (manufacturer: Tongrun; model: VB-1) and evacuated using a vacuum pump (manufacturer: Tingwei; model: TW-4A) to a negative pressure of -70 kPa. This negative pressure was maintained for 10 minutes. The vacuum-sealed reagent trays were then transferred to a cold storage at -20°C. The reagent trays were then assembled into a reagent kit, which was then sealed in a packaging bag using a vacuum packaging machine (manufacturer: Chuangmeng; model: VS600) to a negative pressure of -20 kPa. The package was then frozen and stored in the cold storage for 24 hours. Finally, the reagent trays were 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 held at that negative pressure for 2 hours and 30 minutes.

[0069] The reagent tanks of Examples 1 and 2 were inverted 10 times each to mix them (simulating the mixing treatment of reagents before the actual reaction). After standing for 10 minutes, the temperature and oxygen content of the reagents in the reagent tanks were measured under normal temperature and pressure conditions as the test time was extended (simulating the reaction time during the actual reaction). The oxygen content can be measured by a pH meter. 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 can be understood that when a reagent is placed at a certain ambient temperature for a period of time, the content of gases dissolved in the reagent from the environment and the content of gases released from the reagent into the environment reach a dynamic equilibrium. The oxygen content of the reagent measured at this point is the saturated oxygen content of the reagent. 24℃ is room temperature, and 30℃ and 35℃ are reagent temperatures obtained through a water bath. The measurement results are recorded in Table 2.

[0073] Table 2

[0074]

[0075] Table 1 shows the relationship between the oxygen content of the reagents in Examples 1 and 2 and the test time. The results are shown below. Figure 11 Furthermore, the saturated oxygen content of the reagents measured in Table 2 at 24℃ and 30℃ was added to... Figure 11 In this context, 24℃ is the normal operating temperature for reagent sequencing, while 30℃ can be considered the limiting operating temperature for reagent sequencing. From... 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 of the reagent decreases as the holding time increases. Meanwhile, the oxygen content of the reagents in both Examples 1 and 2 is less than their saturated oxygen content within 20 hours, indicating that a holding time of 10 minutes is sufficient to meet practical requirements.

[0076] Example 3

[0077] The difference from Example 1 is that after the vacuum chamber 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 chamber is evacuated by a vacuum pump, the negative pressure in the reagent tank is -20 kPa.

[0080] Using similar procedures, the temperature and oxygen content changes of the reagents in Examples 3-4 were measured under normal temperature and pressure conditions as the test time increased. The measurement results are recorded in Table 3.

[0081] Table 3

[0082]

[0083] Table 3 shows the relationship between the oxygen content of the reagents in Examples 1 and 3-4 and the test time. The results are shown in Table 3. Figure 12Furthermore, the saturated oxygen content of the reagents measured in Table 2 at 24℃ and 30℃ was added to... Figure 12 In this context, 24℃ is the normal operating temperature for reagent sequencing, while 30℃ can be considered the limiting operating temperature for reagent sequencing. From... Figure 12 It can be seen that the oxygen content of the reagents in Examples 3-4 was less than their saturated oxygen content within 20 hours. At the same time, the oxygen content of the reagents in Examples 3-4 was higher than that of the reagents in Example 1, indicating that the negative pressure condition of -70kPa 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 was not vacuumed after freezing. Instead, the frozen reagent tank was directly sealed and then transferred to a cold storage at -20°C for 24 hours.

[0086] The reagent cells from Example 1 and the comparative example were used for PE150 sequencing on a sequencing platform (manufacturer: BGI; model: DNBSEQ-E25) using a DNBSEQ-E25 chip. The decrease in Q30 (%) for each cycle was then calculated to assess sequencing quality. Q30 is a sequencing quality value. Mathematically, Q30 represents a false positive probability of 0.1%, i.e., an error rate of 0.1%, or an accuracy rate of 99.9%. In sequencing evaluation, to assess the accuracy of reads, Q30 and the proportion of all base quality values ​​greater than Q30 are evaluated. A higher Q30 value reflects better sequencing quality. Specifically, from... Figure 13 and Figure 14 It can be seen that, compared with Example 1, the comparative example showed that due to the entry of air bubbles into the sequencing system, Q30 suddenly dropped by about 3% after a certain cycle and could not be recovered in the later stage, indicating that the reagents in the comparative example affected the sequencing quality due to the precipitation of air bubbles.

[0087] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this application and are not intended to limit it. Although this application has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of this application without departing from the spirit and scope of the technical solutions of this application.

Claims

1. A method for degassing a reagent, characterized in that, Includes the following steps: The reagents are dispensed into reagent tanks and are used in gene sequencing reactions. The reagents include thermal reaction reagents or excision reaction reagents. The reagent tank containing the reagent is subjected to freezing treatment to freeze the reagent into ice; The reagent tank is evacuated to create a first negative pressure, and the pressure is maintained under the first negative pressure to make the gas content in the reagent lower than the gas saturation content of the reagent at room temperature and pressure. The reagent tank is sealed after vacuuming to maintain the first negative pressure inside the reagent tank; The sealed reagent tank is transferred to a storage environment for storage, so that the reagent is kept frozen. In the storage environment, the reagent tank is assembled into a reagent kit, and the reagent kit is packaged in a packaging bag to create a second negative pressure inside the packaging bag, the second negative pressure being equal to or higher than the first negative pressure.

2. The reagent degassing method as described in claim 1, characterized in that, The first negative pressure is between -20 kPa and -70 kPa, and the pressure holding time is not less than 10 minutes.

3. The reagent degassing method as described in claim 2, characterized in that, The first negative pressure is -70 kPa, and the pressure holding time is 10 minutes.

4. The reagent degassing method as described in claim 3, characterized in that, The second negative pressure is -20 kPa.

5. The reagent degassing method as described in claim 1, characterized in that, The temperature of the storage environment is equal to the temperature of the freezing process.

6. The reagent degassing method as described in claim 1, characterized in that, The specific steps of vacuuming the reagent bath include: The frozen reagent tank is placed in a vacuum chamber, so that the inside of the vacuum chamber is connected to the inside of the reagent tank; The vacuum chamber is closed and evacuated to create the first negative pressure inside the reagent tank.

7. The reagent degassing method as described in claim 6, characterized in that, The reagent tank is sealed in a stoppering device, which includes a main body and an electrical control unit electrically connected to the main body. The main body is placed inside the vacuum chamber, and the electrical control unit is placed outside the vacuum chamber. The sealing of the reagent tank specifically includes... The main body is controlled by the electronic control unit to perform corresponding operations, thereby installing the sealing plug into the opening of the reagent tank; Depressurize the vacuum chamber and remove the sealed reagent tank.

8. The reagent degassing method as described in claim 1, characterized in that, After evacuating the reagent bath, the reagent degassing method further includes: The oxygen content in the reagent is measured, and the oxygen content is converted into the gas content of the reagent; The calculated gas content is compared with the gas saturation content of the reagent at room temperature and pressure to determine whether the gas content is lower than the gas saturation content.

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