CN109971830B

CN109971830B - Simple experimental device for DNA spot hybridization

Info

Publication number: CN109971830B
Application number: CN201910269187.8A
Authority: CN
Inventors: 祝建洪; 李阳; 张�雄; 崔蕾
Original assignee: Wenzhou Medical University
Current assignee: Wenzhou Medical University
Priority date: 2019-04-04
Filing date: 2019-04-04
Publication date: 2023-08-18
Anticipated expiration: 2039-04-04
Also published as: CN109971830A

Abstract

The invention discloses a simple experimental device for DNA spot hybridization, which comprises a bearing chassis, wherein one side of the upper end of the bearing chassis is vertically and fixedly connected with a vertical rod, the upper end of the vertical rod is sequentially and movably connected with a first telescopic bracket, a second telescopic bracket and a third telescopic bracket from bottom to top, one end of the third telescopic bracket is connected with a microsyringe needle fixing bracket, two sides of one end surface of the microsyringe needle fixing bracket are uniformly provided with a plurality of mutually corresponding elastic limiting mechanisms, microsyringes needles are arranged between the elastic limiting mechanisms on two sides, one end of the second bracket is connected with an operation table below the microsyringe needle fixing bracket, the upper end surface of the operation table is provided with a nylon membrane, and one side above the bearing chassis is provided with a negative pressure aspirator, and the experimental device has the beneficial effects that: the device has no corresponding limitation on the loading quantity, is simple and easy to operate according to the determination required by experiments, can improve the uncertainty of sample combination caused in the process of natural air drying in manual loading by combining the negative pressure suction device, and saves the time cost.

Description

Simple experimental device for DNA spot hybridization

Technical Field

The invention belongs to the technical field of DNA spot hybridization, and particularly relates to a simple experimental device for DNA spot hybridization.

Background

The DNA spot hybridization technology is a simple experimental technology for combining a DNA sample to be detected on a nitrocellulose membrane or a nylon membrane, preparing a hybridization experiment subsequently, judging whether hybridization and hybridization intensity thereof exist or not through autoradiography, and the experiment can be carried out through a manual sample application or a spot hybridization instrument: the manual sample application is directly performed by using a pipette gun to directly apply the sample on the membrane to wait for the natural air drying of the sample, so that the natural air drying in the manual sample application can cause instability of experimental results and increase experimental time. The number of sample loading holes of a commercial spot hybridization instrument is limited by the instrument, and 48 holes or 96 holes are reserved, so that when the number of samples to be loaded in the experiment does not meet the number of sample loading spaces, the experiment cost can be greatly increased. And meanwhile, the commercial spot hybridization instrument is high in price and high in cost.

Disclosure of Invention

The invention aims to provide a simple experimental device for DNA spot hybridization, which solves the problems at present.

In order to solve the problems, the invention provides a technical scheme that:

the utility model provides a simple and easy experimental apparatus that is used for DNA spot hybridization, includes the bearing chassis, perpendicular fixedly connected with montant in bearing chassis upper end one side, montant upper end from bottom to top swing joint has a flexible support, no. two supports and No. three flexible supports in proper order, no. three flexible supports and all be connected with the bearing between a flexible support and the montant, no. three flexible support one end is connected with microscale pin mount, uniformly be equipped with a plurality of elasticity stop gear that correspond each other on microscale pin mount terminal surface both sides, both sides be equipped with microscale pin between the elasticity stop gear, no. two support one end just is connected with the operation panel below microscale pin mount, the operation panel up end is equipped with the nylon membrane, bearing chassis top one side is equipped with the negative pressure aspirator, be equipped with negative pressure controller on the negative pressure aspirator suction head, the output of negative pressure aspirator passes a flexible support and is connected with the negative pressure aspirator suction head, negative pressure aspirator suction head passes through a flexible support and nylon membrane swing joint.

As the preference, elasticity stop gear comprises fixed block, gag lever post, butt plate, rubber pad, slot, slider, spout, spring pad and extension spring, fixed block and microsyringe needle mount fixed connection just are in microsyringe needle both sides, be equipped with two parallel slots in the fixed block, evenly be connected with a plurality of sliders on the upper and lower wall in the slot, the movable gag lever post that inserts in the slot, be equipped with the spout on the upper and lower both ends of gag lever post, the spout is connected with the slider, the gag lever post passes through slider and spout sliding connection, bottom and gag lever post all are equipped with the spring pad in the one end that the microsyringe needle was kept away from to the gag lever post in the slot, two be connected with extension spring between the spring pad.

Preferably, filter paper is paved on the bottom surface of the nylon membrane.

Preferably, the tip of the suction head is located below the nylon membrane.

The invention has the beneficial effects that:

1. the sample loading hole number of the spot hybridization instrument is limited by the instrument to define 48 holes or 96 holes, the experiment cost is increased when the actual sample loading number of the experiment is not satisfied, the sample loading number of the device is not limited correspondingly, and the device is easy to operate and operate according to the determination required by the experiment.

2. The combined negative pressure aspirator in the device can improve the uncertainty of sample combination caused in the natural air drying process in the manual sample loading process, and simultaneously saves the time cost.

3. Compared with a multi-tube blotting instrument, a hybridization instrument and the like which are sold in the market, the device has lower cost.

Description of the drawings:

for ease of illustration, the invention is described in detail by the following detailed description and the accompanying drawings.

FIG. 1 is a schematic diagram of the structure of the present invention;

FIG. 2 is a schematic view of the elastic limiting mechanism of the present invention;

fig. 3 is an enlarged schematic view of the structure of the present invention at a.

In the figure: 1. a bearing chassis; 2. a vertical rod; 3. a first telescopic bracket; 4. a second bracket; 5. a third telescopic bracket; 6. a bearing; 7. a trace sample injection needle fixing frame; 8. an elastic limit mechanism; 81. a fixed block; 82. a limit rod; 83. a retaining plate; 84. a rubber pad; 85. a slot; 86. a slide block; 87. a chute; 88. a spring pad; 89. a telescopic spring; 9. a microsyringe needle; 10. an operation table; 11. a nylon membrane; 12. a negative pressure aspirator; 13. a negative pressure controller; 14. suction head of negative pressure aspirator.

The specific embodiment is as follows:

as shown in fig. 1-3, the present embodiment adopts the following technical scheme: the utility model provides a simple and easy experimental apparatus that is used for DNA spot hybridization, includes bearing chassis 1, bearing chassis 1 upper end one side vertical fixedly connected with montant 2, montant 2 upper end from bottom to top swing joint has flexible support 3 No. one, no. two supports 4 and No. three flexible supports 5 in proper order, no. three flexible supports 5 one end is connected with microsyringe needle mount 7, uniformly be equipped with a plurality of elasticity stop gear 8 that correspond each other on microsyringe needle mount 7 one end both sides, both sides be equipped with microsyringe needle 9 between the elasticity stop gear 8, no. two supports 4 one end just is connected with operation panel 10 in microsyringe needle mount 7 below, operation panel 10 up end is equipped with nylon membrane 11, bearing chassis 1 top one side is equipped with negative pressure aspirator 12, be equipped with suction head controller 13 on the negative pressure aspirator 12, the output of negative pressure aspirator 12 passes flexible support 3 and is connected with negative pressure aspirator tip 14, negative pressure aspirator 14 passes through flexible support 3 and nylon membrane 11 swing joint.

The elastic limiting mechanism 8 is composed of a fixed block 81, a limiting rod 82, a retaining plate 83, a rubber pad 84, a slot 85, sliding blocks 86, sliding grooves 87, spring pads 88 and telescopic springs 89, wherein the fixed block 81 is fixedly connected with the microsyringe needle fixing frame 7 and is positioned on two sides of the microsyringe needle 9, two parallel slots 85 are arranged in the fixed block 81, a plurality of sliding blocks 86 are uniformly connected on the upper wall and the lower wall in the slot 85, the limiting rod 82 is movably inserted in the slot 85, sliding grooves 87 are formed in the upper end and the lower end of the limiting rod 82, the sliding grooves 87 are connected with the sliding blocks 86, the limiting rod 82 is in sliding connection with the sliding grooves 87 through the sliding blocks 86, the spring pads 88 are respectively arranged at the bottom ends, far away from the microsyringe needle 9, of the limiting rod 82 in the slot 85, and the telescopic springs 89 are connected between the two spring pads 88.

Wherein, filter paper is laid on the bottom surface of the nylon membrane 11, which increases the filtering efficiency of the solvent.

Wherein, the top end of the negative pressure aspirator tip 14 is arranged below the nylon membrane 11, and is matched with the matching of the first telescopic bracket 3 and the bearing 6, the negative pressure aspirator tip can move along with the change of the sample application position, and the matching of the negative pressure aspirator 12 simulates a vacuumized environment, so that DNA in a sample can be combined with the nylon membrane 11 more quickly and better.

The bearing 6 is connected between the third telescopic bracket 5 and the first telescopic bracket 3 and the vertical rod 2, and the first telescopic bracket 3 is matched, so that the sample injection angle of the microscale sample injection needle 9 can be adjusted in all directions, and the manual sample injection of the microscale sample injection needle 9 can be directly simulated.

Specific: when the invention is used, a microsyringe needle 9 (such as 10 mu l, 20 mu l and 100 mu l) with proper volume is selected according to the sample loading volume, the microsyringe needle 9 is placed between the abutting plates 83 of the elastic limiting mechanisms 8 at two sides after the sample is sucked, the microsyringe needle 9 is clamped and fixed on the microsyringe needle fixing frame 7, the nylon membrane 11 and the suction head 14 of the negative pressure suction device 12 are adjusted to determine the area of the sample on the nylon membrane 11, the sample loading volume is manually controlled, the suction intensity of the negative pressure suction device 12 is adjusted through the negative pressure controller 13 to simulate a vacuumizing environment, so that DNA in the sample is combined with the nylon membrane 11 more quickly and better, and the solvent is taken away by the negative pressure suction device 12.

The control mode of the invention is automatically controlled by the negative pressure controller, the control circuit of the negative pressure controller can be realized by simple programming of a person skilled in the art, the supply of power also belongs to the common knowledge in the art, and the invention is mainly used for protecting mechanical devices, so the invention does not explain the control mode and circuit connection in detail.

While the basic principles and main features of the present invention and advantages of the present invention have been shown and described, it will be understood by those skilled in the art that the present invention is not limited by the foregoing embodiments, which are described in the foregoing specification merely illustrate the principles of the present invention, and various changes and modifications may be made therein without departing from the spirit and scope of the invention, which is defined in the appended claims and their equivalents.

Claims

1. A simple experimental device for DNA spot hybridization comprises a bearing chassis (1), and is characterized in that one side of the upper end of the bearing chassis (1) is vertically and fixedly connected with a vertical rod (2), the upper end of the vertical rod (2) is sequentially and movably connected with a first telescopic bracket (3), a second telescopic bracket (4) and a third telescopic bracket (5) from bottom to top, a bearing (6) is connected between the third telescopic bracket (5), the first telescopic bracket (3) and the vertical rod (2), one end of the third telescopic bracket (5) is connected with a microscale sample injection needle fixing frame (7), a plurality of elastic limiting mechanisms (8) which are mutually corresponding are uniformly arranged on two sides of one end surface of the microscale sample injection needle fixing frame (7), microscale sample injection needles (9) are arranged between the elastic limiting mechanisms (8) on two sides, one end of the second telescopic bracket (4) is connected with an operating table (10) below the microscale sample injection needle fixing frame (7), the upper end surface of the operating table (10) is provided with a nylon membrane (11), one side of the upper side of the chassis (1) is provided with a negative pressure device (12), one side of the upper side of the chassis (1) is provided with a negative pressure suction device (12), the negative pressure device (12) is connected with a negative pressure device (12) through the negative pressure device (13) and the negative pressure device (13) is connected with the negative pressure device (13), the negative pressure suction device suction head (14) is movably connected with the nylon membrane (11) through a first telescopic bracket (3), and the top end of the negative pressure suction device suction head (14) is positioned below the nylon membrane (11).

2. The simple experimental apparatus for spot hybridization of DNA according to claim 1, wherein: elastic limit mechanism (8) comprises fixed block (81), gag lever post (82), butt plate (83), rubber mat (84), slot (85), slider (86), spout (87), spring pad (88) and expansion spring (89), fixed block (81) and microsyringe needle mount (7) fixed connection just are in microsyringe needle (9) both sides, be equipped with two parallel slot (85) in fixed block (81), evenly be connected with a plurality of sliders (86) on the upper and lower wall in slot (85), the movable stop lever (82) that has inserted in slot (85), be equipped with spout (87) on both ends about gag lever post (82), spout (87) are connected with slider (86), gag lever post (82) are through slider (86) and spout (87) sliding connection, bottom and gag lever post (82) are kept away from in microsyringe needle (9) one end all are equipped with spring pad (88), two between spring pad (88) flexible spring (89).

3. The simple experimental apparatus for spot hybridization of DNA according to claim 1, wherein: the filter paper is paved on the bottom surface of the nylon membrane (11).