Sponge eraser and nucleic acid removing method

Abstract

The invention provides a sponge eraser and a nucleic acid removing method, and belongs to the technical field of biology. Aiming at pipetting heads with more nucleic acid residues in various pipetting devices, the sponge eraser with a specific structure is designed to be matched with the nucleic acid scavenging agent, so that the nucleic acid scavenging efficiency can be effectively improved, and the negative influence of the nucleic acid residues on downstream experiments is reduced.

Description

Technical Field

[0001] The present application relates to the field of biotechnology, and in particular to a sponge and a nucleic acid removal method. Background Art

[0002] With the continuous development and widespread application of high-throughput sequencing (Next-Generation Sequencing, NGS) technology, the application practice in multiple fields requires higher standards for the automation, stability and efficiency of experimental processes. There are many difficulties in the traditional manual NGS process, such as: cumbersome steps, complex operations, high professional requirements for personnel, low single operation throughput, etc. Automation has become one of the current trends in the development and application of NGS. The automated library construction workstation is a mechanical device that integrates multiple functional modules such as PCR instrument, magnetic rack, temperature control module, pipetting module, oscillation module, etc., to help users realize the automation of NGS processes. Due to the need to process a large number of DNA samples and usually contain PCR amplification processes, the use of the automated library construction workstation will produce more nucleic acid contamination. The accumulation of nucleic acid contamination between multiple batches of experiments can easily lead to the misinterpretation of the final experimental results. Experimental data show that the matrix 96-channel pipetting head is one of the hardest hit areas for the accumulation of nucleic acid contamination. The current common workstation cleaning method is to use 75% anhydrous ethanol and ultrapure water to clean the workstation. For the matrix-type 96-channel pipette head, it may only be simply wiped with 80% anhydrous ethanol or water and dust-free paper. For some pipette heads, the root cannot be wiped, and some users do not even clean the pipette head because the operation is cumbersome. For application scenarios such as tNGS involving multiple rounds of PCR processes, the current method cannot completely remove nucleic acid contamination, which has a significant impact on the accuracy of subsequent experimental results. Summary of the invention

[0003] The purpose of the present application is to provide a sponge and a method for removing nucleic acids. The present application designs a sponge for a 96-channel pipette head, and cooperates with a nucleic acid remover, which is conducive to quickly and efficiently removing the accumulation of nucleic acid contamination remaining on the pipette head and ensuring the reliability of experimental data. The sponge described in the present application can fully ensure that the nucleic acid remover is in contact with the pipette head, avoiding the disadvantage that traditional cleaning methods cannot clean the bottom of the pipette head, while greatly shortening the cleaning time and improving the cleaning efficiency.

[0004] The first aspect of the present application provides a sponge comprising a sponge body and a gasket, wherein the lower end of the sponge body is connected to the upper end of the gasket, a plurality of circular through holes are distributed on the sponge body, and a nucleic acid remover is adsorbed on both the sponge body and the gasket.

[0005] In some embodiments, the nucleic acid remover is an agent that can cause one or more reactions of denaturation, fragmentation, hydrolysis, degradation, precipitation or adsorption of nucleic acid to remove nucleic acid.

[0006] In some embodiments, the nucleic acid is deoxyribonucleic acid (DNA) and / or ribonucleic acid (RNA).

[0007] In some embodiments, the nucleic acid remover includes but is not limited to any nucleic acid remover known to those skilled in the art, such as one or more of ethanol, isopropanol, polyethylene glycol, hydrogen peroxide, peracetic acid, chlorine-containing compounds, sporicides, nucleases, nanometals, strong acids, strong bases, and strong oxidants, and the nuclease includes but is not limited to one or more of ribonucleases (RNase), deoxyribonucleases (DNase), and non-specific nucleases (Non-specific nuclease) known to those skilled in the art.

[0008] In some embodiments, the number and arrangement of the circular through holes match the number and arrangement of the pipette heads, the pipette head is a plug connector connected to the lower end of the pipetting device, used to plug in the pipette tip, the pipette head is usually conical, cylindrical, truncated cone or segmentedly conical and cylindrical or cylindrical and truncated cone protrusions; the pipette tip is a hollow circular tube, the circular tube has a tip opening at the lower end and a plug opening at the upper end, the pipette tip can be connected to the pipette head through the plug opening, the liquid is absorbed into the pipette tip through the tip opening and output from it.

[0009] In some embodiments, the distribution positions of a certain number of the pipetting heads after being arranged in a certain arrangement are the pipetting head holes.

[0010] In some embodiments, the matching means that the number of through holes in the sponge is equal to or greater than the number of the pipetting heads; for example, when a positioning column is also connected to the lower end of the pipetting device, the number of through holes in the sponge is the sum of the number of the pipetting heads and the number of the positioning columns, and at this time, the number of through holes in the sponge is greater than the number of the pipetting heads; for another example, when the number of through holes in the sponge is n times the number of the pipetting heads, so that one sponge can clean n groups of pipetting components, at this time, the number of through holes in the sponge is greater than the number of the pipetting heads.

[0011] In some embodiments, the pipetting heads and the circular through holes are arranged in a matrix, such as a 12×8 matrix.

[0012] In some embodiments, the pipetting head includes but is not limited to any pipetting head known to those skilled in the art, preferably a multi-channel pipetting head, such as an 8-channel pipetting head, a 12-channel pipetting head, a 24-channel pipetting head, and a 96-channel pipetting head.

[0013] In some embodiments, the pipetting device is a pipetting structure capable of realizing a pipetting function, such as a pipetting structure including a pipetting head, a driving element, and a pipetting arm, etc. The pipetting device is not limited to any pipetting device known to a person skilled in the art. The pipetting device is a component used independently or established in an automated instrument, such as an automated pipette, a pipetting workstation, an automated nucleic acid extractor, a high-throughput library construction workstation, etc.

[0014] In some embodiments, the number of the circular through holes is 8, 12, 24 or 96.

[0015] In some embodiments, the diameter of the circular through hole is equal to or slightly larger than or slightly smaller than the diameter of the pipetting head. Here, "slightly" can be understood as a small difference between the diameter of the circular through hole and the diameter of the circular through hole. In the case of the small difference, the circular through hole can completely accommodate the pipetting head, and the outer periphery of the pipetting head contacts the inner periphery of the circular through hole.

[0016] In some embodiments, the diameter of the circular through hole is slightly larger or slightly smaller than the diameter of the pipette tip by up to 10%, for example, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.9%, 8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% and 0.1%.

[0017] In some embodiments, when the diameter of the circular through hole is slightly smaller than the diameter of the pipette head, the maximum elastic deformation of the sponge can ensure that the diameter of the circular through hole is greater than or equal to the diameter of the pipette head, or, the maximum elastic deformation of the sponge can ensure that the circular through hole can completely accommodate the pipette head.

[0018] In some embodiments, the diameter of the pipetting head refers to the average diameter or maximum diameter of a cylinder, cone, or truncated cone comprised by the pipetting head.

[0019] In some embodiments, the circular through hole refers to a cylindrical through hole, the cylindrical through hole penetrates the sponge, and the height of the cylindrical through hole is equal to the height of the sponge.

[0020] In some embodiments, the diameter of the circular through hole is 8-10 mm, including but not limited to 8 mm, 8.1 mm, 8.2 mm, 8.3 mm, 8.4 mm, 8.5 mm, 8.6 mm, 8.7 mm, 8.8 mm, 8.9 mm, 9 mm, 9.1 mm, 9.2 mm, 9.3 mm, 9.4 mm, 9.5 mm, 9.6 mm, 9.7 mm, 9.8 mm, 9.9 mm and 10 mm within the range.

[0021] In some embodiments, the height of the sponge is equal to or slightly less than or slightly greater than the height of the pipetting head. Here, "slightly" can be understood as a small difference between the height of the sponge and the height of the pipetting head, and in the presence of this small difference, the sponge can completely accommodate the pipetting head.

[0022] In some embodiments, the height of the sponge is slightly greater than or less than up to 10% of the height of the pipette tip, for example, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.9%, 8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% and 0.1%.

[0023] In some embodiments, when the height of the sponge is slightly greater than the height of the pipetting head, the maximum elastic deformation of the sponge can ensure that the lower end of the pipetting head touches the gasket.

[0024] In some embodiments, the plurality of circular through holes are arranged in a fixed shape, the length of the sponge and the gasket is greater than or equal to the length of the fixed shape, and the width of the sponge and the gasket is greater than or equal to the width of the fixed shape.

[0025] In some embodiments, when the circular through holes are arranged in an m×n matrix form (m>n), the length of the sponge and the gasket is greater than the sum of the diameters of the m circular through holes, and the width of the sponge and the gasket is greater than the sum of the diameters of the n circular through holes.

[0026] In some embodiments, the length of the sponge is 120-150mm, including but not limited to 120mm, 125mm, 130mm, 135mm, 140mm, 145mm and 150mm within the range; the width of the sponge is 80-120mm, including but not limited to 80mm, 85mm, 90mm, 95mm, 100mm, 105mm, 110mm, 115mm, and 120mm within the range; the width of the sponge is 80-120mm, including but not limited to 80mm, 85mm, 90mm, 95mm, 100mm, 105mm, 110mm, 115mm, and 120mm within the range; the height of the sponge is 8-20mm, including but not limited to 8mm, 9mm, 10mm, 11mm, 12mm, 13mm, 14mm, 15mm, 16mm, 17mm, 18mm, 19mm, and 20mm within the range.

[0027] In some embodiments, the length of the spacer is 120-150 mm, including but not limited to 120 mm, 125 mm, 130 mm, 135 mm, 140 mm, 145 mm, and 150 mm within the range; the width of the spacer is 80-120 mm, including but not limited to 80 mm, 85 mm, 90 mm, 95 mm, 100 mm, 105 mm, 110 mm, 115 mm, and 120 mm within the range; the width of the spacer is 80-120 mm m, including but not limited to 80mm, 85mm, 90mm, 95mm, 100mm, 105mm, 110mm, 115mm, and 120mm within the range; the height of the gasket is 3-20mm, including but not limited to 3mm, 4mm, 5mm, 6mm, 7mm, 8mm, 9mm, 10mm, 11mm, 12mm, 13mm, 14mm, 15mm, 16mm, 17mm, 18mm, 19mm, and 20mm within the range.

[0028] In some embodiments, the length of the gasket is the same as or different from the length of the sponge, preferably, the length of the gasket is greater than or equal to the length of the sponge; the width of the gasket is the same as or different from the width of the sponge, preferably, the width of the gasket is greater than or equal to the width of the sponge; the height of the gasket is the same as or different from the height of the sponge, preferably, the height of the gasket is greater than, less than or equal to the height of the sponge.

[0029] In some embodiments, the sponge and the gasket are integrally formed, or the sponge and the gasket are connected together by any means, such as gluing.

[0030] In some embodiments, the sponge and / or gasket is elastic, and the length, width, height and diameter of the sponge and / or gasket described in this application refer to the length, width, height and diameter in the absence of external force.

[0031] In some embodiments, the sponge is made of a water-absorbing and elastic sponge material, including but not limited to any water-absorbing sponge material known to those skilled in the art, such as polyurethane sponge, polyester sponge and polyvinyl alcohol sponge.

[0032] In some embodiments, the material of the gasket is a hygroscopic material that can absorb water, including but not limited to any sponge, resin, hygroscopic fiber, etc. known to those skilled in the art; preferably, the material of the gasket is a sponge material that can absorb water and has elasticity, including but not limited to polyurethane sponge, polyester sponge and polyvinyl alcohol sponge known to those skilled in the art.

[0033] In some embodiments, the material of the sponge and the material of the gasket are the same or different.

[0034] In some embodiments, the present application exemplarily demonstrates the length, width, height, diameter size and arrangement of the circular through holes of the sponge with a 96-channel pipetting head. When the pipetting head is an 8-channel pipetting head, a 12-channel pipetting head, a 24-channel pipetting head or any specification known to a person skilled in the art, the length, width, height and diameter of the circular through holes of the sponge can correspondingly be any size and arrangement adapted to the aforementioned pipetting head.

[0035] In some embodiments, when the lower end of the pipetting device is connected to other structures besides the pipetting head, for example, when the lower end of the pipetting device is connected to a positioning column, the sponge also includes other through holes with the same shape, quantity and arrangement distribution as the positioning column.

[0036] In some embodiments, the numerical values ​​or numerical ranges of the present application include an error of up to ±20%, up to ±10%, up to ±5%, up to ±4%, up to ±3%, up to ±2% or up to ±1%, where "up to" includes the present number.

[0037] The second aspect of the present application provides a nucleic acid removal method, comprising: using the sponge described in the first aspect, aligning the circular through hole of the sponge with the pipetting head of a pipetting device so that the circular through hole of the sponge body can completely accommodate the pipetting head, and moving the sponge up and down or moving the pipetting head up and down so that the lower end of the pipetting head touches the gasket to remove the nucleic acid residue on the pipetting head;

[0038] The lower end of the pipetting device is provided with a plurality of pipetting heads, and the number and arrangement of the pipetting heads match the number and arrangement of the circular through holes on the sponge.

[0039] In some embodiments, the circular through hole of the sponge body can completely accommodate the pipette head, which means that each of the circular through holes can completely accommodate each of the pipette heads.

[0040] In some embodiments, the distribution positions of a certain number of the pipetting heads after being arranged in a certain arrangement are the pipetting head holes.

[0041] In some embodiments, the circular through hole of the sponge is aligned with the pipetting head of the pipetting device means that the circular through hole of the sponge is aligned with the pipetting head hole of the pipetting device.

[0042] In some embodiments, the matching means that the number of through holes in the sponge is equal to or greater than the number of the pipetting heads; for example, when a positioning column is also connected to the lower end of the pipetting device, the number of through holes in the sponge is the sum of the number of the pipetting heads and the number of the positioning columns, and at this time, the number of through holes in the sponge is greater than the number of the pipetting heads; for another example, when the number of through holes in the sponge is n times the number of the pipetting heads, so that one sponge can clean n groups of pipetting components, at this time, the number of through holes in the sponge is greater than the number of the pipetting heads.

[0043] In some embodiments, the pipetting heads are arranged in a matrix, such as a 12×8 matrix.

[0044] In some embodiments, when the sponge completely accommodates the pipette tip, the lower end of the pipette tip touches or does not touch the upper end of the spacer.

[0045] In some embodiments, the sponge and / or the gasket may be deformed under the action of an external force so that the lower end of the pipetting head touches the upper end of the gasket.

[0046] In some embodiments, the sponge is moved up and down 3-10 times, for example 3-6 times, for example 3 times, 4 times, 5 times or 6 times.

[0047] In some embodiments, the volume of the nucleic acid remover adsorbed on the sponge is, for example, 3-50 mL, including 3 mL, 4 mL, 5 mL, 6 mL, 8 mL, 10 mL, 12 mL, 15 mL, 18 mL, 20 mL, 30 mL, 40 mL and 50 mL within the range.

[0048] In some embodiments, the volume of the nucleic acid remover adsorbed on the sponge is such that the surface of the sponge remains wet and does not exceed the maximum volume that the sponge can adsorb.

[0049] In some embodiments, making the sponge described in the first aspect absorb the nucleic acid remover includes but is not limited to using methods such as spraying, soaking and washing.

[0050] In some embodiments, the method further comprises, after removing nucleic acid residues on the pipette tip, further cleaning the pipette tip using pure water.

[0051] In some embodiments, the further cleaning of the pipette tip with pure water is to clean the nucleic acid remover residue on the pipette tip.

[0052] In some embodiments, the use of pure water to further clean the pipette tip includes: using the sponge described in the first aspect, wherein the nucleic acid remover is replaced with pure water, aligning the circular through hole of the sponge with the pipette head of the pipetting device so that the circular through hole of the sponge can completely accommodate the pipette head, and moving the sponge up and down or moving the pipette head up and down to further clean the pipette head.

[0053] In some embodiments, the volume of pure water adsorbed on the sponge is sufficient to keep the surface of the sponge wet and does not exceed the maximum volume that the sponge can adsorb.

[0054] The third aspect of the present application provides the use of the sponge described in the first aspect in removing nucleic acid contamination.

[0055] In some embodiments, the nucleic acid contamination is nucleic acid contamination on the surface of a pipetting head of a pipetting device, preferably nucleic acid contamination on the surface of a multi-channel pipetting head, preferably nucleic acid contamination at the base of a multi-channel pipetting head. BRIEF DESCRIPTION OF THE DRAWINGS

[0056] Figure 1 : A structural diagram of a pipetting device including a pipetting head;

[0057] Figure 2 : Plan view of the gasket;

[0058] Figure 3 : Plan view of the corpus cavernosum;

[0059] Figure 4 : A three-dimensional diagram of the sponge structure;

[0060] Figure 5 : The position relationship diagram of the pipetting head and the sponge;

[0061] Figure 6 : Example 3: Distribution diagram of the position of the pipette head simulating nucleic acid contamination;

[0062] Figure 7 : Layout of the head, middle and root parts of the pipette tip.

[0063] Specific implementation (embodiment)

[0064] The technical solution of the present application is further explained below in conjunction with the accompanying drawings and through specific implementation methods. However, the following examples are merely simple examples of the present application and do not represent or limit the scope of protection of the present application. The scope of protection of the present application shall be subject to the claims.

[0065] In the following examples, unless otherwise specified, all reagents and consumables used were purchased from conventional reagent manufacturers in the field; unless otherwise specified, all experimental methods and technical means used were conventional methods and means in the field.

[0066] Example 1

[0067] For Vazyme automated high-throughput liquid handling devices (such as the attached Figure 1 , cited from the patent CN219518897U of Novozyme, the exemplary design of the sponge, the pipetting device includes a pipette body and a 96-channel pipetting head, and four positioning columns. The sponge includes Figure 2 The gasket and Figure 3 The sponge shown in the figure has the same length and width, and is connected together by gluing. The sponge includes 12×8 through holes, whose diameter is the same as the diameter of the pipette head, and the spacing between the through holes is the same as the spacing of the pipette head. The height of the sponge is the same as the height of the pipette head. The sponge also includes positioning holes located at the four corners, and the diameter and arrangement of the positioning holes are the same as those of the positioning columns. The complete three-dimensional diagram of the sponge structure is shown in the figure below. Figure 4 shown.

[0068] 50 ng of DNA was smeared at different positions of different pipette tips to simulate nucleic acid contamination. After the DNA was air-dried, a swab was used to wipe the smeared position three times and then a sample was taken. The swab was dissolved in 1000 μl of sterile water, and 9.2 μl was taken after 30 seconds. qPCR detection was performed with Taq Pro Universal SYBR qPCR Master Mix (#Vazyme Q712) as a positive control.

[0069] Use 10mL of Automated Workstation Nucleic Acid Decontamination Agent (#Vazyme VNR11) to spray the sponge until it is wet. The recommended spraying volume should not exceed 20ml. Too much liquid will cause the sponge to be overly wet and drip, affecting the user experience. Use the same method to simulate nucleic acid contamination on the pipette head. Align the wet sponge with the hole of the corresponding pipette head, so that the sponge completely wraps the pipette head to the bottom, so that the tip of the pipette head can touch the gasket, and move up and down 3 times to complete a single cleaning. The positional relationship between the aforementioned sponge and the pipette head during use is as follows: Figure 5 A new sponge was used with ultrapure water for cleaning in the same way, and then a swab was used for sampling, and the sample was tested by qPCR as treatment group 1. The swab sampling method and qPCR test method were the same as above.

[0070] After spraying 10 mL of 75% anhydrous ethanol to wet the surface of the pipette tip, wipe each pipette tip from top to bottom 3 times with dust-free paper, and then clean the pipette tip in the same way using ultrapure water and dust-free paper as routine treatment group 2. The method for simulating nucleic acid contamination of the pipette tip is the same as above.

[0071] A negative control (NTC group, pure water sample) was set up in the experiment, and the clearance efficiency was calculated based on the qPCR results.

[0072] Clearance efficiency = (CT value of treatment group - CT value of positive control group) ÷ (CT value of negative control group - CT value of positive control group) × 100%

[0073] The qPCR test results and removal efficiency of the positive control group, positive control group, conventional treatment group and sponge treatment group are shown in Table 1. After wiping with a sponge and nucleic acid remover VNR11, the removal efficiency can reach 94%, which is much higher than the conventional treatment group, greatly improving the nucleic acid removal effect. In addition, the wiping time is statistically calculated. It takes a total of 6-8 minutes to wipe the pipette head one by one with dust-free paper, and it takes a total of about 5-10 seconds to wipe the entire sponge. This shows that the sponge of the present application can greatly shorten the cleaning time and solve the problem of long time and low efficiency of traditional nucleic acid removal methods.

[0074] Table 1

[0075] Group Average CT value Removal efficiency Positive control group 13.85 / Negative control group 32.88 / VNR11 with sponge 31.87 94.58％ Conventional treatment group (75% anhydrous ethanol) 20.50 34.84％

[0076] Example 2

[0077] As in Example 1, 50 ng of DNA was smeared at different positions of different pipette tips to simulate nucleic acid contamination. After the DNA was air-dried, a sponge was used to wipe with different nucleic acid removers, including VNR11, sodium hypochlorite disinfectant (3% concentration) and sporicide (2% concentration) to conduct the experiment. The specific operation steps were the same as in Example 1. At the same time, the same positive control group, negative control group (NTC) and conventional treatment group (75% anhydrous ethanol) as in Example 1 were set up, and the removal efficiency was calculated by qPCR detection, and the calculation method was the same as in Example 1.

[0078] The qPCR test results and removal efficiency of the positive control group, positive control group, conventional treatment group and three sponge treatment groups are shown in Table 2. The sponge can be used with different nucleic acid removers to achieve efficient removal of nucleic acid contamination in the pipette tip.

[0079] Table 2

[0080] Group Average CT value Removal efficiency Positive control group 12.85 / Negative control group 33.12 / Conventional treatment group (75% anhydrous ethanol) 21.75 43.93％ VNR11 with sponge 31.79 94.56％ Sodium hypochlorite disinfectant (3% concentration) with sponge 30.01 85.69％ Sporicide (2% concentration) with sponge 29.05 80.87％

[0081] Example 3

[0082] Same as Example 1, with reference to Figure 6 The sampling position of the tip head shown in the figure is the head, middle and root of the 12 pipette tips at the four corners and the middle position (such as Figure 7As shown), 50ng DNA was applied to simulate nucleic acid contamination. After the DNA was air-dried, three treatment groups were set up, in which treatment group 1 was cleaned with a sponge and 10mL nucleic acid remover (Vazyme#VNR11), and then cleaned with a sponge and 10mL pure water, and the specific operation steps were the same as in Example 2; treatment group 2 was filled with 10mL nucleic acid remover (Vazyme#VNR11) in a spray pot and sprayed on the surface of the pipette head, until the surface of the pipette head was wet, each pipette head was wiped from top to bottom 3 times with dust-free paper, and then the pipette head was cleaned in the same way with ultrapure water and dust-free paper; treatment group 3 was sprayed with 10mL nucleic acid remover (Vazyme#VNR11) on dust-free paper until it was wet, and each pipette head was wiped from top to bottom 3 times with dust-free paper, and then the pipette head was cleaned with ultrapure water and dust-free paper in the same way; a positive control group and a negative control group (NTC) were set up at the same time. After swabbing the three positions of each group of pipette heads, the removal efficiency was calculated by qPCR detection. The calculation method was the same as in Example 1. The proportions of the number of positions with a removal efficiency greater than 90%, less than 80%, and greater than 80% and less than 90% in each treatment group were counted respectively.

[0083] The results of the removal efficiency ratios of the three treatment methods of VNR11, namely sponge wiping, spraying and dust-free paper, are shown in Table 3. VNR11 combined with a sponge can ensure that different positions of the pipette head can be cleaned more efficiently. For example, for the middle part of the pipette head that is more difficult to clean, the removal efficiency of all sponge test groups can be above 90%. For the most difficult to clean root of the pipette head, the two conventional methods of spraying or dust-free paper with nucleic acid remover wiping have a removal efficiency of 16.67% and 8.33% of the test groups respectively below 80%. The use of the sponge of the present application with a nucleic acid remover can make the removal efficiency of all test groups reach more than 80%, of which 91.67% of the test groups can achieve a nucleic acid removal efficiency of more than 90%, which is significantly higher than the conventional control group. Compared with traditional methods, the sponge of the present application can improve the comprehensiveness of nucleic acid removal, solving the problem that traditional methods cannot clean the root of the pipette head.

[0084] Table 3

[0085]

[0086] Example 4

[0087] The nucleic acid removal test of the pipette tip was carried out using a sporicide with a sponge, spraying and dust-free paper, and the specific experimental operation steps and data processing method were the same as those in Example 3. The number ratios of positions with a removal efficiency greater than 90%, less than 80%, and greater than 80% and less than 90% in each treatment group were counted respectively. The results of the removal efficiency ratios of VNR11 with a sponge, spraying and dust-free paper are shown in Table 4.

[0088] As shown in Table 4, the experimental data after replacing other types of nucleic acid removers are consistent with the trend of Example 3. The sponge wipe combined with the sporicide can still improve the nucleic acid removal efficiency for the pipette head, especially the root of the pipette head which is more difficult to clean. The removal efficiency of the sponge wipe test group all reached more than 80%, while the spray and dust-free paper combined with the sporicide as the control had a removal rate of 25% of the test groups below 80%. Combined with the results of Example 3, it can be seen that the sponge wipe combined with different types of nucleic acid removers can achieve a comprehensive nucleic acid removal effect, reduce operation time, and improve nucleic acid removal efficiency.

[0089] Table 4

[0090]

Claims

1. A sponge, comprising a sponge body and a gasket, wherein the lower end of the sponge body is connected to the upper end of the gasket, a plurality of circular through holes are distributed on the sponge body, and a nucleic acid remover is adsorbed on both the sponge body and the gasket.

2. The sponge according to claim 1, wherein the nucleic acid remover is one or more of ethanol, isopropanol, polyethylene glycol, hydrogen peroxide, peracetic acid, chlorine-containing compounds, sporicides, nucleases, nanometals, strong acids, strong bases and strong oxidants.

3. The sponge according to claim 1, wherein the number and arrangement of the circular through holes match the number and arrangement of the pipetting heads, and the pipetting head is a plug connector arranged at the lower end of the pipetting device, and the plug connector is used to plug the pipetting tip; preferably, the pipetting head is a multi-channel pipetting head, preferably an 8-channel pipetting head, a 12-channel pipetting head, a 24-channel pipetting head or a 96-channel pipetting head; preferably, the number of the circular through holes is 8, 12, 24 or 96; preferably, the circular through holes are arranged in a 12×8 matrix form.

4. The sponge according to claim 3, wherein the diameter of the circular through hole is equal to, slightly larger than, or slightly smaller than the diameter of the pipetting head; preferably, the diameter of the circular through hole is 8-10 mm.

5. The sponge according to claim 1, wherein the plurality of circular through holes are arranged in a fixed shape, the length of the sponge and the gasket is greater than or equal to the length of the fixed shape, and the width of the sponge and the gasket is greater than or equal to the width of the fixed shape; Optionally, when the circular through holes are arranged in an m×n matrix form (m>n), the length of the sponge and the gasket is greater than the sum of the diameters of the m circular through holes, and the width of the sponge and the gasket is greater than the sum of the diameters of the n circular through holes; Optionally, the length of the sponge is 120-150 mm, the width of the sponge is 80-120 mm, the length of the gasket is 120-150 mm, and the width of the gasket is 80-120 mm.

6. The sponge wipe according to claim 3, wherein the height of the sponge body is equal to, slightly smaller than, or slightly larger than the height of the pipetting head; preferably, the height of the sponge body is 8-20 mm.

7. The sponge wipe according to claim 3, wherein the pipetting device is a pipetting structure capable of realizing a pipetting function; optionally, the pipetting device is a component used independently or established in an automated instrument; preferably, the pipetting device is an automated pipette, an automated pipetting workstation, an automated nucleic acid extractor or a high-throughput library construction workstation.

8. The sponge wiper according to claim 1, wherein the sponge body is made of a water-absorbing and elastic sponge material; preferably, the sponge material is one or more of polyurethane sponge, polyester sponge and polyvinyl alcohol sponge; Optionally, the gasket is made of a hygroscopic material that can absorb water; optionally, the hygroscopic material is a sponge, resin or hygroscopic fiber; preferably, the gasket is made of a sponge material that can absorb water and has elasticity; preferably, the sponge material is one or more of a polyurethane sponge, a polyester sponge and a polyvinyl alcohol sponge.

9. A method for removing nucleic acid, comprising: Using the sponge according to any one of claims 1 to 8, aligning the circular through hole of the sponge with the pipetting head of a pipetting device so that the circular through hole of the sponge body can completely accommodate the pipetting head, moving the sponge up and down or moving the pipetting head up and down so that the lower end of the pipetting head touches the gasket to remove the nucleic acid residue on the pipetting head; The lower end of the pipetting device is provided with a plurality of pipetting heads, and the number and arrangement of the pipetting heads match the number and arrangement of the circular through holes on the sponge.

10. The method of claim 9, wherein when the sponge completely accommodates the pipetting head, the lower end of the pipetting head touches or does not touch the upper end of the gasket; Optionally, the number of times of moving the sponge up and down or moving the pipetting head up and down is 3-10 times; Optionally, the sponge and / or the gasket may be deformed under the action of an external force so that the lower end of the pipetting head touches the upper end of the gasket.

11. The method of claim 9, further comprising, after removing the nucleic acid residue on the pipette tip, further cleaning the pipette tip with pure water; Optionally, use pure water to further clean the nucleic acid remover residue on the pipette tip; Optionally, the further cleaning of the pipetting tip with pure water comprises: Use the sponge described in claim 1, wherein the nucleic acid remover is replaced with pure water, align the circular through hole of the sponge with the pipetting head of the pipetting device so that the circular through hole of the sponge can completely accommodate the pipetting head, and move the sponge up and down or move the pipetting head up and down to further clean the pipetting head.

12. Use of the sponge according to any one of claims 1 to 8 in removing nucleic acid contamination.

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