CN207727071U - A kind of ethyl alcohol remove device for nucleic acid extraction experiment - Google Patents

Abstract

The utility model discloses an ethanol removal device for nucleic acid extraction experiment, which comprises a fan main body, a test tube box and a box cover capable of being buckled on the test tube box, the test tube box is equipped with an EP tube frame, and the EP tube frame is A number of test tube holes are arranged, and EP tubes containing nucleic acids to be dried can be placed in the test tube holes. The main body of the fan is provided with a fan and a power supply for the fan. The main body of the fan is provided with a fan port. The air outlet is located in front of the fan, and an air nozzle is installed on the air outlet, and the other end of the air nozzle is provided with a horizontal flat nozzle; the left and right sides of the box cover are respectively provided with an air inlet and an air outlet. The utility model has changed the previous way of drying the test tubes by vertical wind, using the negative pressure effect generated by the horizontal wind to realize the rapid volatilization of residual ethanol in the EP test tubes, effectively reducing the steps and time for removing residual ethanol in the batch nucleic acid extraction process, and reducing the extraction time. Probability of sample contamination.

Description

An ethanol removal device for nucleic acid extraction experiments

technical field

The utility model relates to the technical field of nucleic acid extraction devices, in particular to an ethanol removal device used for nucleic acid extraction experiments.

Background technique

Nucleic acid precipitation is required during the nucleic acid extraction process, and in order to ensure the quality of the nucleic acid, it is necessary to wash the nucleic acid precipitation with ethanol and remove the salt and water. Therefore, after washing the nucleic acid, the washed precipitate must be dried to facilitate the dissolution of the nucleic acid precipitate in the later stage. In the existing operation, the generally followed steps are: use 1ml of 75% ethanol to wash the precipitate twice, then use 1ml of absolute ethanol to wash the precipitate, and pour out the absolute ethanol in the test tube after centrifugation; then use a centrifuge to centrifuge the test tube , use a pipette gun to absorb the ethanol at the bottom of the precipitation; use the method of inverting the test tube on the test bench to let it drain naturally, or use the sterile wind in the ultra-clean workbench to promote the volatilization of the remaining ethanol, but the speed of natural air drying is too slow, super The clean workbench is restricted by the vertical wind direction, which often leads to pollutants falling into the test tube, causing nucleic acid contamination and degradation, and bringing worries to the follow-up high-precision molecular tests.

Contents of the invention

In order to solve the deficiencies and defects of the prior art, the purpose of this utility model is to provide an ethanol removal device for nucleic acid extraction experiments, which has a fast ethanol removal speed and will not pollute the nucleic acid samples in the test tube.

In order to achieve the above object, the technical solution of the present utility model is an ethanol removal device for nucleic acid extraction experiments, including a blower main body, a test tube box and a box cover that can be buckled on the test tube box, and the EP tube rack is housed in the test tube box , the EP tube frame is arranged with some test tube holes, the EP tubes containing the nucleic acid to be dried can be placed in the test tube holes, the fan and the power supply for the fan are arranged in the main body of the fan, and the main body of the fan is equipped with An air outlet is provided, and the air outlet is located directly in front of the fan. An air nozzle is installed on the air outlet, and the other end of the air nozzle is provided with a horizontal flat nozzle; the left and right sides of the box cover are respectively equipped with The air outlet and the air outlet, the flat nozzle can be inserted from the air inlet, the flat nozzle, the air inlet, and the air outlet are located on the same horizontal line, and the airflow changes brought about by the rotation of the fan form a horizontal wind along the fan outlet, the air nozzle, and the air inlet. It passes through the top of the EP pipe rack and blows out from the air outlet, thereby forming a negative pressure above the EP pipe.

Preferably, a filter is provided inside the flat mouth, and a replaceable filter membrane for filtering air is provided on the side of the filter close to the fan, and the periphery of the filter is closely attached to the filter through an elastic sealing ring. online.

Preferably, the filter screen is a metal screen, and the metal screen is a stainless steel screen or a copper-nickel alloy screen.

Preferably, the air inlet is located above the nozzle of the EP pipe, and the air inlet is also provided with a flat air inlet, and the flat nozzle can be inserted from the flat air inlet.

Preferably, the air nozzle is installed on the air outlet in a plug-in manner.

Preferably, the main body of the fan is also provided with a wind speed adjustment knob for adjusting the wind speed, and the wind speed adjustment knob is electrically connected to the fan through a controller.

Preferably, the power supply is a built-in power supply, and the built-in power supply includes a battery box, a battery cover, and a battery placed in the battery box.

Preferably, the power supply is an external power supply, and the external power supply includes an external wire and an external plug electrically connected to the fan.

Preferably, a counterweight is provided at the bottom of the main body of the fan; the rear end of the main body of the fan is tightly fixed on the side wall of the ultra-clean workbench through a suction cup.

Preferably, several hasp rods are movably connected to the outer wall of the test tube box, the hasp rods are located below the air inlet, and the main body of the fan is provided with a fixing rod corresponding to the position of the hasp rods. The rod can be hung on the fixed rod.

Preferably, the fixing rod is C-shaped, and the other end of the buckle rod is provided with an inverted U-shaped hook that can hang on the fixing rod.

Preferably, the left and right sides of the main body of the fan are hingedly connected with an L-shaped connecting rod, and the ends of the L-shaped connecting rod are provided with suction cup nozzles that can be adsorbed on the outer walls of the front and rear ends of the test tube box.

Preferably, the utility model also includes a grid plate, the top of the grid plate is provided with grid holes, the grid plate is located above the EP pipe, and the aperture of the grid hole is larger than that of the EP pipe nozzle. aperture, the nozzle of the EP pipe is exposed in the mesh hole, and the entire top cover of the EP pipe is pressed under the mesh plate.

Preferably, the utility model also includes a grid plate, the top of the grid plate is provided with grid holes, the grid plate is located above the EP pipe, and the aperture of the grid hole is larger than that of the EP pipe nozzle. The aperture, the nozzle of the EP tube and the top cover of the EP tube are all exposed in the grid holes, and the ears of the top cover are pressed under the grid plate.

A handle is provided on the top of the grid plate to facilitate the placement of the grid plate.

The utility model also provides a method for using the ethanol removal device, comprising the following steps:

(1) Soak the elastic sealing ring, grid plate, air nozzle with filter screen, and box cover in DEPC water, sterilize under high temperature and high pressure, and dry in an oven to remove RNase;

(2) After putting the filter membrane into the air nozzle, insert the sealing ring so that the filter membrane is tightly attached to the filter screen, install the air nozzle at the air outlet, and make the flat mouth of the air nozzle lie in the horizontal direction;

(3) Put the EP tube rack into the test tube box, insert the EP tube containing the nucleic acid containing ethanol washing residue into the test tube hole of the EP tube rack, make the nozzle of the EP tube open, and put the grid plate , put the lid on the test tube box;

(4) Insert the flat nozzle into the air inlet, and then fix the test tube box and the main body of the fan together;

(5) Turn on the power and run the fan at a wind speed of 13-52m/s for 10min-20min to complete the removal of ethanol.

The utility model method has the following advantages:

The utility model has changed the previous way of drying the test tubes by vertical wind, using the negative pressure effect generated by the horizontal wind to realize the rapid volatilization of residual ethanol in the EP test tubes, effectively reducing the steps and time for removing residual ethanol in the batch nucleic acid extraction process, and at the same time reducing the extraction time. Probability of sample contamination.

Description of drawings

Fig. 1 is the structural representation of the utility model;

Fig. 2 is the structural representation of the lid of the utility model;

Fig. 3 is the fixing mode of test tube box in embodiment 1;

Fig. 4 is the fixing mode of test tube box in embodiment 3;

Figure 5 is a top view of Figure 4;

Fig. 6 is the fixing mode of test tube box in embodiment 4;

Fig. 7 is the structural representation of the grid plate of embodiment 5;

Fig. 8 is the structural representation of the grid plate of embodiment 6;

Fig. 9 is the effect diagram of RNA electrophoresis obtained by drying using the ethanol removal device of the present invention;

Reference signs: 1. Fan main body; 2. Test tube box; 3. EP tube rack; 4. Test tube hole; 5. EP tube; 51. Nozzle; 52. Top cover; 521. Cover ears; 6. Fan; 7 , power supply; 8, fan outlet; 9, air nozzle; 10, flat nozzle; 11, filter screen; 12, filter membrane; 13, box cover; 14, elastic sealing ring; 15, air inlet; 16, air outlet; 17 , flat air inlet nozzle; 18, counterweight block; 19, suction cup; 20, hasp rod; 21, fixed rod; 22, inverted U-shaped hook; 23, L-shaped connecting rod; 24, suction cup nozzle; 25, net Grid plate; 26, grid holes; 27, handle; 28, wind speed adjustment knob.

Detailed ways

The following examples are used to illustrate the utility model, but not to limit the scope of the utility model.

Example 1

As shown in Figures 1-3, an ethanol removal device for nucleic acid extraction experiments includes a blower main body 1, a test tube box 2 and a box cover 13 that can be buckled on the test tube box 2, and the test tube box 2 is equipped with EP tubes Rack 3. Several test tube holes 4 are arranged on the EP tube rack 3. EP tubes 5 containing nucleic acids to be dried are inserted in the test tube holes 4. The specification of the EP tubes 5 is 1.5-2.0ml. The main body of the fan 1 is provided with a fan 6 and a power supply 7 for powering the fan 6. The power supply 7 is a built-in power supply, and the built-in power supply includes a battery box, a battery cover and a battery placed in the battery box.

The fan main body 1 is provided with a fan outlet 8, the fan outlet 8 is located directly in front of the fan 6, and the fan outlet 8 is equipped with a tuyere 9, and the tuyere 9 can be installed on the fan by plugging and unplugging. On the mouth 8, therefore, the utility model can take off the tuyere 9 separately to carry out the treatment of sterilization and RNase removal. The fan main body 1 is also provided with a wind speed adjusting knob 28 for adjusting the wind speed, and the wind speed adjusting knob 28 is electrically connected with the fan 6 through the controller.

The other end of the air nozzle 9 is provided with a horizontal flat nozzle 10, which can converge the airflow generated by the rotation of the fan 6 and form a horizontal wind. The flat nozzle 10 is provided with a filter screen 11, and the filter screen 11 is provided with a replaceable filter membrane 12 for filtering air on the side close to the fan 6, and the periphery of the filter membrane 12 passes through an elastic sealing ring 14 Closely attached to the filter screen 11. The filter screen 11 is a metal mesh, and the metal mesh is a stainless steel mesh or a copper-nickel alloy mesh. The setting of the filter membrane 12 can ensure that the wind entering the test tube box 2 does not contain impurities such as bacteria, RNA, RNase, dust, etc. The setting of the filter screen can provide mechanical support for the filter membrane 12, and the elastic sealing ring 14 makes the filter membrane 12 more stable. .

The left and right sides of the box cover 13 are respectively provided with an air inlet 15 and an air outlet 16 , and the flat nozzle 10 can be inserted through the air inlet 15 . In order to ensure that the connection between the flat nozzle 10 and the box cover 13 is more stable and air-tight, the utility model is also provided with a flat air inlet nozzle 17 on the air inlet 15, and the flat nozzle 10 can be inserted from the flat inlet nozzle 17. The flat nozzle 10, the air inlet 15, and the air outlet 16 are located on the same horizontal line, and the airflow changes caused by the rotation of the fan 6 form a horizontal wind along the fan mouth 8, the air nozzle 9, and the air inlet 15 and enter the top of the EP pipe frame 3 and flow from Air outlet 16 is blown out, thereby forms negative pressure above EP tube 5, accelerates the evaporation of ethanol in the test tube. And the incoming wind is sterile, RNA-free, and has no risk of RNase contamination, and will not blow directly to the wall of the EP tube or the nucleic acid sample in the EP tube, thus effectively avoiding sample (DNA or RNA) contamination.

And, described air inlet 15 is positioned at the nozzle 51 top of described EP pipe 5, thereby guarantees that the horizontal wind that enters from air inlet 15 can not directly act on test tube, when avoiding air-flow passing between test tube box 2 and lid 13 cause the test tube to shake

The bottom of the fan main body 1 is also provided with a counterweight 18, and the rear end of the fan main body 1 is tightly fixed on the side wall in the ultra-clean workbench through a suction cup 19. The setting of the counterweight and the suction cup 19 ensures During the operation of the fan 6, the fan main body 1 is firmly fixed and does not move. The outer wall of the test tube box 2 is connected with some snap bars 20, and the snap bars 20 can be connected with the test tube box by hinged connection (such as ball hinge connection) or other connection modes (also can be fixed connection mode). connect. The snap rod 20 is located below the air inlet 15, and the main body of the fan 1 is provided with a fixed rod 21 corresponding to the position of the snap rod 20, and the snap rod 20 can be attached to the fixed rod 21, thereby avoiding The test tube box 2 is blown and displaced during use.

Example 2

Different from Embodiment 1, the power supply 7 is an external power supply, and the external power supply includes an external wire and an external plug electrically connected to the fan 6 .

Example 3

The difference from Embodiment 1 is that, as shown in FIGS. 4-5 , the fixing rod 21 is C-shaped, and the other end of the snap rod 20 is provided with an inverted U-shaped hook 22 that can hang on the fixing rod 21 . Therefore, in the blowing process, the snap rod 20 is not forced to shake left and right along the fixed rod 21, and the design of the inverted U-shaped snap hook 22 also restricts the front and rear displacement of the snap rod 20. Therefore, the design of this embodiment makes The fixing effect of the kit is better and more reliable.

Example 4

The difference from Embodiments 1 and 4 is that, as shown in FIG. 6 , the left and right sides of the fan main body 1 are hingedly connected (such as a ball hinge connection) with an L-shaped connecting rod 23 , and the L-shaped connecting rod 23 The end is provided with the sucker mouth 24 that can be adsorbed on the outer walls of the front and rear ends of the test tube box 2 .

Example 5

Different from Embodiment 1, as shown in Figure 7, the utility model also includes a grid plate 25, and a grid hole 26 is opened on the top of the grid plate 25, and the grid plate 25 is located in the EP pipe 5 Above, the aperture of the mesh hole 26 is greater than the aperture of the nozzle 51 of the EP pipe 5, the nozzle 51 of the EP pipe 5 is exposed in the mesh hole 26, and the entire top cover 52 of the EP pipe 5 is pressed Under the grid plate 25, the setting of the grid plate 25 is beneficial to fix the position of the EP pipe 5, thereby preventing the EP pipe 5 from shaking due to the horizontal wind at its top.

A handle 27 is provided on the top of the grid plate 25 to facilitate the placement of the grid plate 25 .

Example 6

Different from Embodiment 5, as shown in Figure 8, the utility model also includes a grid plate 25, and a grid hole 26 is opened on the top of the grid plate 25, and the grid plate 25 is located on the EP pipe 5 Above, the aperture of the mesh hole 26 is greater than the aperture of the nozzle 51 of the EP pipe 5, the nozzle 51 of the EP pipe 5 and the top cover 52 of the EP pipe 5 are all exposed in the mesh hole 26, the EP The cover ear 521 of the top cover 52 of the tube 5 is pressed under the grid plate 25. Compared with the embodiment 5, the setting of the grid plate 25 in this embodiment not only helps to fix the position of the EP tube 5, but also prevents the EP tube from 5 because the horizontal wind at its top shakes, and the grid plate 25 does not touch the interface of the top cover 52 that is fastened with the nozzle 51 of the EP tube 5, thereby further avoiding sample contamination.

A handle 27 is provided on the top of the grid plate 25 to facilitate the placement of the grid plate 25 .

Example 7

The utility model also provides the use method of the ethanol removal device of embodiment 1-6, comprising the following steps:

(1) Soak the elastic sealing ring 14, the grid plate 25, the air nozzle 9 with the filter screen 11, and the box cover 13 in DEPC water, sterilize under high temperature and high pressure, and dry in an oven to remove RNase.

(2) After putting the filter membrane 12 into the air nozzle 9, insert the elastic sealing ring 14 so that the filter membrane 12 is tightly attached to the filter screen, install the air nozzle 9 at the fan outlet 8, and make the air nozzle 9 flat The mouth 10 is located in a horizontal direction.

(3) EP tube frame 3 is put into test tube box 2, the EP tube 5 that will fill the nucleic acid that contains ethanol washing raffinate is inserted on the test tube hole 4 of EP tube frame 3, makes the nozzle 51 of EP tube 5 be in opening state, cover the box cover 13 on the test tube box 2.

(4) Insert the flat nozzle 10 into the air inlet 15, and then fix the test tube box 2 and the blower main body 1 together.

(5) Turn on the power supply 7 to run the fan 6 at 52m/s for 10min to complete the removal of ethanol.

In order to extract more clean and pollution-free samples, the above ethanol removal process can be completed in the ultra-clean workbench. Utilize the above method to use the device of the present utility model in a short period of time, there is no need to replace the filter membrane 12 and the sterilization of the box cover 13, the grid plate 25, the air nozzle 9, and the sterilization of the test tube box 2 and the EP tube rack 3 does not need to be frequent conduct. Therefore, when using continuously for a short period of time, it is only necessary to insert the EP tube 5 filled with ethanol-washed nucleic acid into the test tube hole 4, and then cover the lid 13 and connect it with the flat mouth 10. After the nucleic acid is precipitated, centrifuged, and ethanol is discarded, the utility model directly carries out negative pressure drying, and does not need time-consuming and labor-intensive operations such as centrifuging again and extracting residual ethanol with a pipette gun, and also avoids sample pollution, especially for large-scale nucleic acid extraction. has great significance.

1. Utilize the ethanol removal device of the present utility model to dry RNA

Wash the precipitate with 1ml of absolute ethanol, and pour out the absolute ethanol in the test tube after centrifugation; then centrifuge the test tube with a centrifuge, and use a pipette gun to absorb the ethanol at the bottom of the precipitate; Drain, or blow the ultra-clean workbench to promote the volatilization of remaining ethanol. The ethanol-cleared RNA sample was subjected to electrophoresis using the device and method of the present invention. The electrophoresis result is shown in FIG. 9 , and the two RNA bands are clear without obvious degradation.

2. Air-drying speed of ethanol under different drying methods

The dehydrated alcohol that adds 1ml is washed DNA precipitation, ethanol is poured out after centrifugation, then directly utilizes natural drying respectively, ultra-clean typhoon (8m/s), the ethanol device of the present utility model (adjustment wind speed is respectively 13m/s) , 26m/s, 39m/s, 52m/s) for drying, each treatment is provided with three repetitions, the results are shown in Table 1 (the premise of Table 1 comparison is under the same situation of ethanol). Table 1 shows that if the format is changed, it will be messed up. There is no obvious difference between the horizontal wind and the vertical wind.

Table 1 Air-drying speed comparison of ethanol under different drying methods

wind speed(m/s) Dry naturally(0) Ultra-clean table air dry(8) 8 13 26 39 52 Evaporation speed (μL/min) 0.25 0.5 0.5 1.0 1.4 1.7 2.0

As shown in Table 1, at the same wind speed, vertical wind still has an advantage. The advantage of horizontal wind is that it is less likely to cause pollution.

3. The time spent in different drying methods

Add 1 ml of 75% ethanol to wash the DNA precipitate twice, then add 1 ml of absolute ethanol to wash the DNA precipitate, prepare three groups (48 samples in each group), and each group has 3 repetitions. The first group of samples and the second group of samples are processed conventionally, centrifuged twice, and the remaining part of ethanol is extracted with a pipette, then the first group of samples is dried naturally, and the second group of samples is dried with ultra-clean bench air (8m/s ) was air-dried; the third group did not need to carry out secondary centrifugation and the extraction of the remaining part of ethanol, and directly utilized the device of the present utility model (adjusting the wind speed to be 52m/s) to carry out DNA drying. The results are shown in Table 2.

Table 2 The removal time of ethanol under different operating modes

wind speed(m/s) Dry naturally(0) Ultra-clean table air dry(8) The utility model (52) time-consuming (min) 80±10min 60±10min 20±5min

As shown in Table 2, because the operation steps of residual ethanol centrifugation and secondary extraction are saved, the utility model greatly saves the removal time of ethanol and improves the efficiency of nucleic acid extraction.

Although the utility model has been described in detail with general description and specific examples above, some modifications or improvements can be made on the basis of the utility model, which will be obvious to those skilled in the art. Therefore, the modifications or improvements made on the basis of not departing from the spirit of the utility model all belong to the protection scope of the utility model.

Claims (9)

1. a kind of ethyl alcohol remove device for nucleic acid extraction experiment, it is characterised in that：Including fan main body, test tube case and energy The box cover on test tube case is enough withheld, EP pipe supports are housed in the test tube case, several test tube holes, institute are placed on the EP pipe supports The EP for filling nucleic acid to be dried pipes can be placed by stating in test tube hole；Equipped with fan and be fan power supply in the fan main body Power supply, the fan main body are equipped with wind turbine mouth, and the wind turbine mouth is located at the front of fan, wind is equipped on the wind turbine mouth The other end of mouth, the tuyere is equipped with horizontal flat mouth；The box cover is respectively arranged on the left side and the right side air intake vent and air outlet, The flat mouth can be inserted into from air intake vent, and the flat mouth, air intake vent, air outlet are located at same horizontal line, the fan rotation The air-flow variation brought forms horizontal wind above EP pipe supports and from air outlet blowout along wind turbine mouth, tuyere, air intake vent, thus Negative pressure is formed above EP pipes.

2. the ethyl alcohol remove device according to claim 1 for nucleic acid extraction experiment, it is characterised in that：The flat mouth Interior to be equipped with filter screen, the filter screen is being equipped with replaceable filter membrane for filtering air, the filter close to the side of fan It is tightly attached on filter screen by elastic seal ring on the periphery of film.

3. the ethyl alcohol remove device according to claim 2 for nucleic acid extraction experiment, it is characterised in that：The air intake vent Above the nozzle of the EP pipes, be additionally provided on the air intake vent it is flat enter tuyere, the flat mouth can from it is flat enter tuyere Interior insertion；The tuyere is mounted on by way of plug on wind turbine mouth.

4. the ethyl alcohol remove device according to claim 3 for nucleic acid extraction experiment, it is characterised in that：The wind turbine master The wind speed adjusting knob for adjusting wind speed is additionally provided on body, the wind speed adjusting knob is electrically connected by controller and fan It connects；The bottom of the fan main body is additionally provided with clump weight；The rear end of the fan main body is tightly secured in ultra-clean work by sucker Make on the side wall in platform.

5. the ethyl alcohol remove device according to claim 4 for nucleic acid extraction experiment, it is characterised in that：The test tube case Outer wall on be connected with several hasp bars, the hasp bar is located at the lower section of air intake vent, the fan main body be equipped with The corresponding fixed link in hasp bar position, the hasp bar can be affiliated in fixed link.

6. the ethyl alcohol remove device according to claim 5 for nucleic acid extraction experiment, it is characterised in that：The fixed link For c-type, the other end of the hasp bar is equipped with the inverted U that can be affiliated in fixed link and has contact with.

7. the ethyl alcohol remove device according to claim 4 for nucleic acid extraction experiment, it is characterised in that：The wind turbine master A L-type connecting rod is hingedly connected at left and right sides of body, the end of the L-type connecting rod is equipped with before can be adsorbed on test tube case Sucker mouth on the outer wall of both ends afterwards.

8. the ethyl alcohol remove device according to claim 4 for nucleic acid extraction experiment, it is characterised in that：It further include a net Panel offers grid hole above the waffle slab, and the waffle slab is located at the top of EP pipes, the aperture of the grid hole More than the aperture of EP pipe nozzles, the nozzle of the EP pipes is exposed in grid hole, and the entire head cover of the EP pipes is pressed on grid Under plate.

9. the ethyl alcohol remove device according to claim 4 for nucleic acid extraction experiment, it is characterised in that：It further include a net Panel offers grid hole above the waffle slab, and the waffle slab is located at the top of EP pipes, and the aperture of the grid hole is big In the aperture of EP pipe nozzles, the nozzle of the EP pipes and the head cover of EP pipes are exposed in grid hole, the lid ear of the head cover Place is pressed under waffle slab.