CN102134552B - Nucleic acid hybridization method and nucleic acid hybridization instrument utilizing same - Google Patents
Nucleic acid hybridization method and nucleic acid hybridization instrument utilizing same Download PDFInfo
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- CN102134552B CN102134552B CN201010620523.8A CN201010620523A CN102134552B CN 102134552 B CN102134552 B CN 102134552B CN 201010620523 A CN201010620523 A CN 201010620523A CN 102134552 B CN102134552 B CN 102134552B
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- hybridization
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- groove
- nucleic acid
- vibrating trough
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- 238000007899 nucleic acid hybridization Methods 0.000 title claims abstract description 37
- 238000000034 method Methods 0.000 title description 11
- 238000009396 hybridization Methods 0.000 claims abstract description 73
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 42
- 239000007788 liquid Substances 0.000 claims abstract description 41
- 238000010438 heat treatment Methods 0.000 claims abstract description 11
- 239000012528 membrane Substances 0.000 claims abstract description 8
- 239000002699 waste material Substances 0.000 claims description 28
- 230000002572 peristaltic effect Effects 0.000 claims description 7
- 238000005070 sampling Methods 0.000 claims description 5
- 230000007306 turnover Effects 0.000 claims description 3
- 239000011435 rock Substances 0.000 claims description 2
- 238000002474 experimental method Methods 0.000 abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 4
- 238000005213 imbibition Methods 0.000 description 3
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000010808 liquid waste Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000003805 vibration mixing Methods 0.000 description 1
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Abstract
The invention discloses a nucleic acid hybridization instrument comprising a reagent bottle, a liquid inlet device, a liquid-transferring device, a vibrating trough device and a control system, wherein the liquid inlet device is connected with the liquid-transferring device and provided with a micropump and a membrane pump, and the micropump and the membrane pump are used for driving reagents to flow into the liquid-transferring device from the reagent bottle; the liquid-transferring device is arranged parallel to the vibrating trough device and provided with a needle tube device, the vibrating trough is provided with a hybridization trough, and the needle tube device is used for adding the reagents into the corresponding hybridization trough for hybridization; and the control system is connected with the liquid inlet device, the liquid-transferring device and the vibrating trough device through a controller. The nucleic acid hybridization instrument can carry out full-automatic experiments so as to save a great deal of time, and adopts the liquid inlet device and the liquid-transferring device for sucking and feeding liquid so as to achieve high speed and convenient operation; the vibrating trough device vibrates automatically, and a heating plate is used for carrying out constant temperature heating so that heating can be more direct and fast; and the control device is used for controlling operation so as to improve the accuracy of the experiment and reduce experimental errors.
Description
Technical field
The present invention relates to a kind of nucleic acid hybridization and utilize the nucleic acid hybridization oven of the method.
Background technology
At present, general nucleic acid hybridization is all to adopt the mode of imbibition manually and liquid feeding to carry out nucleic acid hybridization, and the liquid of mixing is placed in water-bath and is heated and vibration mixing in water-bath, adopt and carry out in this way nucleic acid hybridization, get liquid measure levels of precision be difficult to guarantee, manual operations to spend the more time particularly need to have plurality of reagents participate in time, the heating in water bath time is long, has greatly extended whole experimental period.
Summary of the invention
The technical problem to be solved in the present invention be to provide a kind of automatically, a kind of nucleic acid hybridization and utilize the nucleic acid hybridization oven of the method easily.
In order to solve the problems of the technologies described above, the technical solution adopted for the present invention to solve the technical problems is: a kind of nucleic acid hybridization is provided, it is characterized in that, comprise the following steps:
1) clean running: carry out first equipment being cleaned before dot blot experiment, in clear water access arrangement, each liquid line and hybridization groove clean, and after cleaning, waste liquid are taken away;
2) fill pipe running: each reagent is admitted to corresponding liquid line, and is full of after each liquid line, and unnecessary liquid discharge is collected;
3) dress film running: by vibrating trough ejecting device, place film bar in corresponding hybridization groove in vibrating trough, then vibrating trough is pushed back to device specified location;
4) hybridization running: different reagent is flowed in each hybridization groove by setting order by liquid line, carries out preheating at hybridization groove, shakes up, hybridization, coupling, and hybridization finishes to take out the colour developing of film bar;
5) clean running: experiment finishes rear repeating step 1), device is cleaned.
Wherein, step 1) with step 2) in clear water and each reagent complete feed liquor process by surge pump and micro pump acting in conjunction.
Wherein, step 3) in described crossover process, comprise hybridization groove is carried out to preheating and heating, and hybridization groove is vibrated.
For solving the problems of the technologies described above, another technical solution used in the present invention is: a kind of nucleic acid hybridization oven that utilizes aforesaid method is provided, comprises reagent bottle, feeding device, liquid-transfering device, vibrating trough device and Controlling System;
Described feeding device is connected with liquid-transfering device, and described feeding device is provided with micro pump and surge pump, and described micro pump and surge pump drive reagent to enter liquid-transfering device by reagent bottle;
Described liquid-transfering device and vibrating trough device be arranged in parallel, on described liquid-transfering device, establish needle tube device and the rectilinear orbit that needle tube device is moved left and right and move up and down, described vibrating trough device is provided with hybridization groove, and described needle tube device adds corresponding hybridization groove to hybridize reagent;
In described Controlling System, connect feeding device, liquid-transfering device and vibrating trough device by controller.
Wherein, the needle tube device on described liquid-transfering device is double needle, and two syringe needles be arranged in parallel, and the spacing between the spacing between described double needle and hybridization groove equates.
Wherein, described syringe needle comprises sample needle for adding reagent and for extracting the sampling pin of waste liquid out.
Wherein, further on hybridization instrument, be provided with tooth bar, the gear of described tooth bar and vibrating trough device matches to merge and drives vibrating trough device turnover hybridization instrument, and described Controlling System is provided with group of motors, described group of motors band carry-over bar motion.
Wherein, hybridization groove in described vibrating trough device is level or the length direction setting that tilts at hybridization groove with respect to the horizontal plane, in the time that hybridization groove is horizontally disposed with, one end in hybridization slot length direction is provided with groove, in the time that hybridization groove is obliquely installed, hybridization groove with respect to the horizontal plane lower one end is provided with groove, is provided with gear pin or fence for locating Hybond membrane between Hybond membrane and groove.
Wherein, in described vibrating trough device, be provided with for the hot-plate to the heating of hybridization groove with for driving the vibrating device of hybridization groove vibration.
Wherein, nucleic acid hybridization oven of the present invention further comprises waste collecting device, described waste collecting device comprises waste collection bottle and peristaltic pump, described waste collection bottle connects each liquid line of each nucleic acid hybridization oven, and described peristaltic pump drives the waste liquid in waste collection bottle to discharge nucleic acid hybridization oven to special sewer line.
The invention has the beneficial effects as follows, in can setting by this nucleic acid hybridization, automatically test, save a large amount of time, adopt feed liquor and liquid-transfering device to carry out imbibition and liquid feeding, speed is quick, easy to operate, carries out automatic vibration, hot-plate thermostatically heating by vibrating trough, heats more direct, faster, handle and greatly improved experiment tolerance range by control device control, reduce experimental error.
Accompanying drawing explanation
Below in conjunction with accompanying drawing, the specific embodiment of the present invention is described in further detail.
Fig. 1 is the perspective view in nucleic acid hybridization oven embodiment of the present invention;
Fig. 2 is the vertical view in nucleic acid hybridization oven embodiment of the present invention;
Fig. 3 is the front view in nucleic acid hybridization oven embodiment of the present invention;
Wherein, 1: reagent bottle;
2: feeding device; 21: micro pump; 22: surge pump;
3: liquid-transfering device; 31: needle tube device;
4: vibrating trough device; 41: hybridization groove; 42: tooth bar;
5: Controlling System; 51: group of motors;
6: waste collecting device; 61: waste collection bottle; 62: peristaltic pump.
Embodiment
By describing technology contents of the present invention, structural attitude in detail, being realized object and effect, below in conjunction with embodiment and coordinate accompanying drawing to be explained in detail.
Please participate in the lump Fig. 1, Fig. 2 and Fig. 3, the embodiment of nucleic acid hybridization oven of the present invention comprises reagent bottle 1, feeding device 2, liquid-transfering device 3, vibrating trough device 4 and Controlling System 5;
Described reagent bottle 1 can arrange multiple reagent bottles as required, and gives each reagent bottle and necessarily number and place different experiments reagent, for example, be designed to four large reagent bottle A, B, C, D and three little reagent bottle E, F, G, and reagent bottle D places clear water;
Described feeding device 2 is connected with liquid-transfering device 3, described feeding device 2 is provided with micro pump 21 and surge pump 22, micro pump 21 is in proper order for unit control to liquid, and surge pump 22 is to control with the time, described micro pump 21 and surge pump 22 actings in conjunction drive reagent to enter liquid-transfering device 3 by reagent bottle 1;
Described liquid-transfering device 3 be arranged in parallel with vibrating trough device 4, liquid-transfering device 3 is matched in experimentation mutually with vibrating trough device 4, described liquid-transfering device 3 is provided with the needle tube device 31 and the rectilinear orbit that needle tube device is moved left and right and move up and down for liquid feeding, by this rectilinear orbit, needle tube device 31 can carry out up and down liquid feeding and get liquid, liquid is moved in left and right, described vibrating trough device 4 is provided with hybridization groove 41, and described needle tube device 31 adds reagent in corresponding hybridization groove 41 and carries out hybrid experiment;
In described Controlling System 5, connect feeding device 2, liquid-transfering device 3 and vibrating trough device 4 by controller;
Described accounting hybridization instrument is provided with tooth bar 42, and described tooth bar 42 matches to merge with the gear of vibrating trough device 4 and drives 4 turnover of vibrating trough device to adjust hybridization instrument, and described Controlling System 5 is provided with group of motors 51, and described group of motors 51 is with carry-over bar 42 to move.
While carrying out nucleic acid hybridization, comprise following step:
1) clean running: carry out first equipment being cleaned before dot blot experiment, Controlling System 5 is by controller control feeding device 2, clear water is sent into the each liquid line of equipment and hybridization groove 41 from reagent bottle D by micro pump 21 and surge pump 22, each liquid line and hybridization groove 41 are cleaned, clean up rear waste liquid and take away by the needle tube device 31 in liquid-transfering device 3;
2) fill pipe running: Controlling System 5 is by controller control feeding device 2, from different reagent bottle 1, corresponding reagent is sent into corresponding liquid line by micro pump 21 and surge pump 22, fill the liquid that previous section in pipe is mixed with clear water and discharge, reagent is full of to each liquid line;
3) dress mould running: by control device 5 Vibration on Start-up groove ejecting devices, vibrating trough device 4 is released to hybridization instrument, open the cover plate on hybridization groove 41, experimental film bar is placed in the interior corresponding hybridization groove 41 of vibrating trough device 4, cover the cover plate on hybridization groove 41, vibrating trough device 4 is pushed back to hybridization instrument specified location;
4) hybridization running: experimental in Controlling System 5 different hybridization programs need to be set, disparity items is having little difference aspect time and reagent dosage, instrument reaches higher accuracy within the specific limits, controlling micro pump 21 and surge pump 22 drives different reagent to flow in each hybridization groove 41, carry out preheating, shake up, hybridization and coupling, hybridization finishes rear taking-up film bar colour developing.
5) clean running: experiment finishes rear repeating step 1), device is cleaned.
In aforesaid method, in described vibrating trough device 4, be provided with the hot-plate for heating to hybridization groove 41 and the vibrating device for driving hybridization groove 41 to vibrate, a side below hybridization groove 41 is provided with a snib structure, be provided with a motor in a corresponding side, motor is provided with eccentric wheel, trace one end connects snib, and one end connecting eccentric wheel, has formed eccentric straight-line running gear thus.When driven by motor eccentric wheel rotates, can drive hybridization groove 41 front and back to rock back and forth by the linear slide of the slide block below eccentric wheel, connecting rod, snib and base plate and guide rail composition.
Step 3) in described crossover process, by hot-plate, the liquid in hybridization groove 41 is carried out to preheating and heating, vibrate to the liquid in hybridization groove 41 by vibrating device, described hybridization groove 41 is level or the length direction setting that tilts at hybridization groove 41 with respect to the horizontal plane, in the time that hybridization groove 41 is horizontally disposed with, one end on hybridization groove 41 length directions is provided with groove, in the time that hybridization groove 41 is obliquely installed, hybridization groove 41 with respect to the horizontal plane lower one end is provided with groove, is provided with gear pin or fence for locating Hybond membrane between Hybond membrane and groove.
Adopt above-mentioned nucleic acid hybridization oven and nucleic acid hybridization, can in setting program, automatically test, save a large amount of experimental periods, adopt feed liquor and liquid-transfering device to carry out imbibition and liquid feeding, speed is quick, easy to operate, carries out automatic vibration, hot-plate thermostatically heating by vibrating trough, heats more direct, faster, handle and greatly improved experiment tolerance range by control device control, reduce experimental error.
In the present embodiment, needle tube device 31 on described liquid-transfering device 3 is double needle, two syringe needles be arranged in parallel, spacing between spacing between described double needle and hybridization groove 41 equates, two syringe needles can, with a kind of reagent of fashionable dress, when experiment is carried out, carry out liquid feeding and get liquid two connected hybridization grooves 41 simultaneously, improve the speed that reagent adds experiment, reduce experimental period.
The syringe needle of described needle tube device 31 comprises sample needle 33 for adding reagent and for extracting the sampling pin 34 of waste liquid out, can add experiment reagent to hybridization groove by sample needle 33, take the waste liquid in hybridization groove away with sampling pin 34 equally, sample needle 33 and sampling pin 34 are combined, and the extraction that fast and easy is carried out experiment reagent adds.
In the present embodiment, nucleic acid hybridization oven also further comprises waste collecting device 6, described waste collecting device 6 comprises waste collection bottle 61 and peristaltic pump 62, described waste collection bottle 61 connects each liquid line of each nucleic acid hybridization oven, and described peristaltic pump 62 drives the waste liquid in waste collection bottle 61 to discharge nucleic acid hybridization oven to special sewer line.
By waste collecting device 6 is set, conveniently process the liquid waste producing in experiment, by relevant device, waste collection got up and enter together special sewer line, avoid reagent directly to enter sewer line, produce pollution and destruction to environment.
The foregoing is only embodiments of the invention; not thereby limit the scope of the claims of the present invention; every equivalent structure or conversion of equivalent flow process that utilizes specification sheets of the present invention and accompanying drawing content to do; or be directly or indirectly used in other relevant technical fields, be all in like manner included in scope of patent protection of the present invention.
Claims (6)
1. a nucleic acid hybridization oven, is characterized in that, comprises reagent bottle, feeding device, liquid-transfering device, vibrating trough device and Controlling System;
Described feeding device is connected with liquid-transfering device, and described feeding device is provided with micro pump and surge pump, and described micro pump and surge pump drive reagent to enter liquid-transfering device by reagent bottle;
Described liquid-transfering device and vibrating trough device be arranged in parallel, described liquid-transfering device is provided with needle tube device and the rectilinear orbit that needle tube device is moved left and right and move up and down, described vibrating trough device is provided with hybridization groove, and described needle tube device adds corresponding hybridization groove to hybridize reagent;
One side of described hybridization groove below is provided with a snib structure, be provided with a motor in a corresponding side, described motor is provided with eccentric wheel, connect snib by trace one end, one end connecting eccentric wheel, when driven by motor eccentric wheel rotates, rock back and forth before and after driving hybridization groove by the linear slide of the slide block below eccentric wheel, connecting rod, snib and base plate and guide rail composition;
In described Controlling System, connect feeding device, liquid-transfering device and vibrating trough device by controller;
Further on hybridization instrument, be provided with tooth bar, the gear of described tooth bar and vibrating trough device matches to merge and drives vibrating trough device turnover hybridization instrument, and described Controlling System is provided with group of motors, described group of motors band carry-over bar motion.
2. nucleic acid hybridization oven according to claim 1, is characterized in that, the needle tube device on described liquid-transfering device is double needle, and two syringe needles be arranged in parallel, and the spacing between the spacing between described double needle and hybridization groove equates.
3. nucleic acid hybridization oven according to claim 2, is characterized in that, described syringe needle comprises sample needle for adding reagent and for extracting the sampling pin of waste liquid out.
4. nucleic acid hybridization oven according to claim 1, hybridization groove in described vibrating trough device is level or the length direction setting that tilts at hybridization groove with respect to the horizontal plane, in the time that hybridization groove is horizontally disposed with, one end in hybridization slot length direction is provided with groove, in the time that hybridization groove is obliquely installed, hybridization groove with respect to the horizontal plane lower one end is provided with groove, is provided with gear pin or fence for locating Hybond membrane between Hybond membrane and groove.
5. nucleic acid hybridization oven according to claim 4, is characterized in that, is provided with for the hot-plate to the heating of hybridization groove with for driving the vibrating device of hybridization groove vibration in described vibrating trough device.
6. according to the nucleic acid hybridization oven described in claim 1 to 5 any one, it is characterized in that, further comprise waste collecting device, described waste collecting device comprises waste collection bottle and peristaltic pump, described waste collection bottle connects each liquid line of each nucleic acid hybridization oven, and described peristaltic pump drives the waste liquid in waste collection bottle to discharge nucleic acid hybridization oven to special sewer line.
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| CN201010620523.8A CN102134552B (en) | 2010-12-31 | 2010-12-31 | Nucleic acid hybridization method and nucleic acid hybridization instrument utilizing same |
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| CN201010620523.8A CN102134552B (en) | 2010-12-31 | 2010-12-31 | Nucleic acid hybridization method and nucleic acid hybridization instrument utilizing same |
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| CN102134552B true CN102134552B (en) | 2014-05-21 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102899248B (en) * | 2012-10-25 | 2014-11-26 | 杭州奥盛仪器有限公司 | Humidification temperature-controlled oscillation type hybridization device |
| CN105176790B (en) * | 2014-05-27 | 2018-09-25 | 昆明寰基生物芯片产业有限公司 | Microarray hybridization elution instrument |
| CN105936873B (en) * | 2016-06-13 | 2018-06-22 | 珠海赛乐奇生物技术股份有限公司 | A kind of cleaning structure of hybridization instrument |
| CN107629952B (en) * | 2017-09-18 | 2024-03-19 | 益善生物技术股份有限公司 | Nucleic acid hybridization instrument |
| CN111254044A (en) * | 2018-12-01 | 2020-06-09 | 艾康生物技术(杭州)有限公司 | Reaction cup, molecular hybridization instrument, application of molecular hybridization instrument and detection method |
| CN109576150A (en) * | 2018-12-29 | 2019-04-05 | 西南民族大学 | Membrane DNA chip hybridization instrument |
| CN112300911A (en) * | 2020-10-27 | 2021-02-02 | 广州和实生物技术有限公司 | Nucleic acid detector and nucleic acid detection method |
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| JP2003093039A (en) * | 2001-09-21 | 2003-04-02 | Juki Corp | Hybridization apparatus and method for detecting nucleic acid in sample by using the same |
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| CN2582367Y (en) * | 2002-12-13 | 2003-10-29 | 武汉中敏基因工程有限公司 | Gene proliferation hybridization apparatus |
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