CN105273976A - Nuclease inactivating instrument with diethylpyrocarbonate isolating function and application of nuclease inactivating instrument - Google Patents
Nuclease inactivating instrument with diethylpyrocarbonate isolating function and application of nuclease inactivating instrument Download PDFInfo
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- CN105273976A CN105273976A CN201510684453.5A CN201510684453A CN105273976A CN 105273976 A CN105273976 A CN 105273976A CN 201510684453 A CN201510684453 A CN 201510684453A CN 105273976 A CN105273976 A CN 105273976A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/99—Enzyme inactivation by chemical treatment
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/02—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using physical phenomena
- A61L2/04—Heat
- A61L2/06—Hot gas
- A61L2/07—Steam
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Abstract
The invention provides a nuclease inactivating instrument with a diethyl pyrocarbonate isolating function and application of the nuclease inactivating instrument. The nuclease inactivating instrument structurally comprises a DEPC (diethylpyrocarbonate) dilution and reserve tank (1), an autoclaves sterilizer (2), a connection valve (5), motors (6), an air pump (7), a DEPC well (8), a water inlet valve (9), a reserve tank water inlet hopper (10), a mixing blade (11), a tip box rack (12), a drain valve (13), an exhaust valve (14), a top cover (15) of the autoclaves sterilizer, a heating pipe (16), a centrifugal drum (17), an EP pipe fixing chamber (21) and the like. By the aid of the nuclease inactivating instrument, a great amount of tips and EP pipes can be subjected to RNase inactivation for one time, and accordingly workload is greatly relieved for laboratory technicians. Due to DEPC recycling, chemical reagents are saved; since DEPC water is heated for decomposition prior to being discharged, environment protection is realized.
Description
Technical field
Patent of the present invention relates to molecular biology experiment instrument field, specifically possesses nuclease deactivation instrument and the application thereof of diethylpyrocarbonate isolation features.
Background technology
RNA extraction purification is the groundwork content of molecular biology research, is also the primary work of transcriptional level research.Due to RNA lytic enzyme (RNase) extensive existence and be difficult to the obstinate person's character of deactivation, make the extracting and purifying of RNA and follow-up work difficult especially.Although Trizol reagent and the widespread use of all kinds of RNA extracting and purifying test kit make RNA extracting and purifying become relative simple, the work relating to RNA remains in molecular biology research the problem allowing people have a headache most.After all, the subject matter of RNA work is the pollution preventing RNase.RNase is ubiquitous, and in any step of experimental implementation, any accidental carelessness or ill-considered operation all likely cause the pollution of RNase, thus cause the failure of whole experiment.Therefore, strict Control release condition, avoids any possible RNase to pollute, and is the key ensureing Success in Experiment.
The instrument EP pipe directly contacted with RNA sample and the RNase deactivation of rifle head are extremely strict, high-temperature sterilization twice after general employing diethylpyrocarbonate (DEPC) aqueous solution soaking, concrete operations are as follows: one, EP pipe and rifle head 0.1%DEPC aqueous solution immersion treatment 12 hours at 37 DEG C.Two, twice autoclaving 30 minutes further deactivation RNas at 120 DEG C the residual DEPC of high temperature removing.
The following three place's problems of existence in RNase deactivation prior art: cause DEPC water to be difficult to enter rifle head and EP pipe inside because of rifle head and EP canal float on the DEPC water surface or because rifle head and EP inner air tube cannot get rid of when one, soaking rifle head and EP pipe, do not reach the effect of RNase deactivation.Two, during immersion rifle head, rifle head is scattered in container, needs rifle head to put into rifle head box one by one during sterilizing.Rifle head substantial amounts, and DEPC has severe toxicity, in order to laboratory technician is safe and do not pollute the rifle head soaked, usually uses tweezers gripping rifle head one by one, also needs to throw away DEPC water in rifle head, and such laboratory technician's working strength is very large, and the risk having DEPC poisoning.Three, traditional method is not soaked DEPC water and can not be reused under the environment of fully-closed, generally directly pours water drain into, causes reagent waste and cause environmental pollution.
Summary of the invention
Patent of the present invention provides a kind of efficient and possess the RNase deactivation instrument of DEPC isolation features, its structure comprises: DEPC dilution and conservation tank (1) (hereinafter referred to as conservation tank), high-pressure sterilizing pot (2), base (3), universal wheel (4), connection valve (5), electric motor (6), air pump (7), DEPC well (8), water intaking valve (9), conservation tank influent hopper (10), paddle (11), rifle head carriage (12), water discharge valve (13), vent valve (14), Autoclave top cover (15), heating tube (16), centrifugal barrel (17), rifle head carriage snap close (18), rifle head (19), EP pipe fixed bin baffle plate (20), EP pipe fixed bin (21), rifle first gear plate (22), rifle head box panel (23), high-pressure sterilizing pot influent hopper (25), wherein the function of air pump can be replaced by water pump (24).
Conservation tank and high-pressure sterilizing pot are arranged on the base of band universal wheel, convenient mobile.Universal wheel has snap close, avoids sliding when not moving.Connection valve is arranged on the below of conservation tank and high-pressure sterilizing pot, and control DEPC water flows between conservation tank and high-pressure sterilizing pot.Whole instrument has that three electric motor are respectively paddle (6-1), air pump (6-2), centrifugal barrel (6-3) provide power.Air pump electric motor rotate and reverse lower realization to conservation tank pressurization and bleed.DEPC well is connected on water inlet pipe, has screw capping, can seal.Centrifugal barrel is arranged on high-pressure sterilizing pot inside, bottom connects electric motor, dismountable rifle head carriage becoming 60-85 degree inclination angle with centrifugal barrel is installed in centrifugal barrel, rifle head (19) is placed on the rifle head box panel (23) that rifle head box carries upper (only direct after RNase deactivation panel is put back to rifle head box), panel can be fixed by rifle head carriage, and pins outlet (18); Be provided with band above rifle head carriage and permitted lacunary rifle first gear plate (22), shape is as fallen " recessed " font, and can block rifle head and float in DEPC water, the aperture of metal sheet can allow the unrestricted flow of DEPC water enter rifle head.Rifle head carriage is uniformly distributed in centrifugal barrel, balances during to enable centrifugal; Rifle head carriage arranges 5-15 layer in centrifugal barrel, and every layer 4-10, each rifle head carriage can place 1-3 rifle head box panel.Can put into EP pipe fixed bin (21) after rifle head carriage dismounting in centrifugal barrel in centrifugal barrel, the baffle plate (20) be fixed in centrifugal barrel is placed in top, and when avoiding centrifugal, EP pipe moves; In order to balance time centrifugal, EP pipe fixed bin is arranged to the large spaces such as 4 or 8.Above structure is made by stainless material except electric motor, base and universal wheel.
Possess and make DEPC water between conservation tank and high-pressure sterilizing pot, flow the structure of function except air pump is to conservation tank pressurization with except bleeding, also have a kind of alternative structure to be mounted in water pump on conservation tank and autoclaving pot bottom pipe connecting, water pump electric motor (6-4) rotate and reverse drive under realization DEPC water is flowed back and forth between conservation tank and high-pressure sterilizing pot.
The present invention possesses the nuclease deactivation instrument of diethylpyrocarbonate isolation features and applies in use first according to DEPC water concentration and conservation tank volume computing DEPC consumption, tightens lid after a certain amount of DEPC is added DEPC well.Open water intaking valve, from influent hopper, add a certain amount of distilled water, DEPC is poured conservation tank by distilled water.Open paddle electric motor, DEPC is uniformly dissolved.EP pipe is evenly put into each space of EP pipe fixed bin, cover baffle plate.Rifle head carriage is installed, and the rifle head box panel of the head that carries a gun is fixed in rifle head carriage, pin outlet, and close high-pressure sterilizing pot top cover.Open connection valve, and open air pump electric machine conservation tank is pressurizeed, make DEPC water all enter high-pressure sterilizing pot, and flood centrifugal barrel.Close air pump electric machine and connection valve, open centrifugal barrel electric motor and control at 10-100r/min by rotating speed, under rotation situation, DEPC water fully enters in rifle head and EP pipe, closes centrifugal barrel electric motor, soaks 30min.Open the gentle pump motor of connection valve after abundant immersion, air pump is bled to conservation tank, makes DEPC water get back to conservation tank, leaves a small amount of liquid as autoclaved steam source (or open top cover add distilled water as autoclaved steam source).Opening centrifugal barrel electric motor carries out centrifugal, is thrown away by DEPC water in rifle head and EP pipe.Regulate high-pressure sterilizing pot air pressure and time, autoclaving twice, again open centrifugal barrel electric motor and carry out centrifugal, the water of condensation produced in rifle head during sterilizing and EP pipe is thrown away.DEPC water cycle uses repeatedly, is pumped into by DEPC water after heating in high-pressure sterilizing pot makes DEPC decompose and opens water discharge valve discharge.
The invention has the beneficial effects as follows: the present invention once can carry out RNase deactivation to a large amount of rifle head and EP pipe, significantly reduces laboratory technician's workload.This instrument, to the management of DEPC water seal, avoids laboratory technician to contact the generation of DEPC poisoning.This instrument recycles DEPC and has saved chemical reagent, and discharges after DEPC water heating and decomposition treatment, protects environment.
Accompanying drawing explanation
Fig. 1 is nuclease deactivation instrument and the application longitudinal section structural map thereof that the present invention possesses diethylpyrocarbonate isolation features.
Fig. 2 is nuclease deactivation instrument and the application centrifugal barrel longitudinal section structural map thereof that the present invention possesses diethylpyrocarbonate isolation features.
Fig. 3 is nuclease deactivation instrument and the signal of application rifle head carriage vertical section structure thereof that the present invention possesses diethylpyrocarbonate isolation features.
Fig. 4 be the present invention possess diethylpyrocarbonate isolation features nuclease deactivation instrument and application the rifle head carriage right side look meaning.
Fig. 5 is nuclease deactivation instrument and the application EP pipe fixed bin vertical view thereof that the present invention possesses diethylpyrocarbonate isolation features.
Fig. 6 is nuclease deactivation instrument and the application water pump structure schematic diagram thereof that the present invention possesses diethylpyrocarbonate isolation features.
Fig. 7 is that the present invention possesses the nuclease deactivation instrument of diethylpyrocarbonate isolation features and applying high voltage Autoclave influent hopper installation site thereof and structural representation.
Embodiment
Embodiment one:
As accompanying drawing 1: patent of the present invention provides a kind of efficient and possess the RNase deactivation instrument of DEPC isolation features, and its structure comprises: DEPC dilution and conservation tank (1) (hereinafter referred to as conservation tank), high-pressure sterilizing pot (2), base (3), universal wheel (4), connection valve (5), electric motor (6), air pump (7), DEPC well (8), water intaking valve (9), influent hopper (10), paddle (11), rifle head carriage (12), water discharge valve (13), vent valve (14), Autoclave top cover (15), heating tube (16), centrifugal barrel (17).Conservation tank and high-pressure sterilizing pot are arranged on the base of band universal wheel, convenient mobile.Universal wheel has snap close, avoids sliding when not moving.Connection valve is arranged on the below of conservation tank and high-pressure sterilizing pot, and control DEPC water flows between conservation tank and high-pressure sterilizing pot.Whole instrument has that three electric motor are respectively paddle (6-1), air pump (6-2), centrifugal barrel (6-3) provide power.Air pump electric motor rotate and reverse lower realization to conservation tank pressurization and bleed.DEPC well is connected on water inlet pipe, has screw capping, can seal.
As accompanying drawing 3 and accompanying drawing 4: centrifugal barrel is arranged on high-pressure sterilizing pot inside, bottom connects electric motor, dismountable rifle head carriage becoming 70-85 degree inclination angle with centrifugal barrel is installed in centrifugal barrel, rifle head (19) is placed on the narrow meshed panel of rifle head box (23), panel can be fixed by rifle head carriage, and pins outlet (18); Be provided with band above rifle head carriage and permitted lacunary metal sheet, rifle head can be blocked and float in DEPC water, and the unrestricted flow of DEPC water can be allowed to enter rifle head.
As accompanying drawing 2 and accompanying drawing 5: can put into EP pipe fixed bin (21) after the rifle head carriage dismounting in centrifugal barrel, top is placed and is fixed on centrifugal barrel Internal baffle (20), and when avoiding centrifugal, EP pipe moves; In order to balance time centrifugal, EP pipe fixed bin is arranged to the large spaces such as 4 or 8.Above structure is made by stainless material except electric motor, base and universal wheel.
The present invention possesses the nuclease deactivation instrument of diethylpyrocarbonate isolation features and applies in use first according to DEPC water concentration and conservation tank volume computing DEPC consumption, tightens lid after a certain amount of DEPC is added DEPC well.Open water intaking valve, from influent hopper, add a certain amount of distilled water, DEPC is poured conservation tank by distilled water.Open paddle electric motor, DEPC is uniformly dissolved.EP pipe is evenly put into each space of EP pipe fixed bin, cover baffle plate.Rifle head carriage is installed, and the rifle head box panel of the head that carries a gun is fixed in rifle head carriage, pin outlet, and close high-pressure sterilizing pot top cover.Open connection valve, and open air pump electric machine conservation tank is pressurizeed, make DEPC water all enter high-pressure sterilizing pot, and flood centrifugal barrel.Close air pump electric machine and connection valve, opening centrifugal barrel motor control rotating speed is 20r/min, and under rotation situation, DEPC water fully enters in rifle head and EP pipe, closes centrifugal barrel electric motor, soaks 30min.Open the gentle pump motor of connection valve after abundant immersion, air pump is bled to conservation tank, makes DEPC water get back to conservation tank, leaves a small amount of liquid as autoclaved steam source (or open top cover add distilled water as autoclaved steam source).Opening centrifugal barrel electric motor carries out centrifugal, is thrown away by DEPC water in rifle head and EP pipe.Regulate high-pressure sterilizing pot air pressure and time, autoclaving twice, again open centrifugal barrel electric motor and carry out centrifugal, the water of condensation produced in rifle head during sterilizing and EP pipe is thrown away.DEPC water cycle uses repeatedly, is pumped into by DEPC water after heating in high-pressure sterilizing pot makes DEPC decompose and opens water discharge valve discharge.
Embodiment two:
As attached 1,6 and 7: tighten lid after a certain amount of DEPC is added DEPC well.Open water intaking valve, from influent hopper, add a certain amount of distilled water, DEPC is poured conservation tank by distilled water.Open paddle electric motor, DEPC is uniformly dissolved.EP pipe is evenly put into each space of EP pipe fixed bin, cover baffle plate.Rifle head carriage is installed, and the rifle head box panel of the head that carries a gun is fixed in rifle head carriage, pin outlet, and close high-pressure sterilizing pot top cover.Open connection valve (5), and open water pump motor, make DEPC water all enter high-pressure sterilizing pot, and flood centrifugal barrel.Switch off the pump electric motor and connection valve, and open centrifugal barrel electric motor and control at 10-100r/min by rotating speed, under rotation situation, DEPC water fully enters in rifle head and EP pipe, closes centrifugal barrel electric motor, soaks 30min.Open connection valve and pump motor after abundant immersion, pump motor reverses, and makes DEPC water get back to conservation tank, leaves a small amount of liquid as autoclaved steam source (or adding distilled water from high-pressure sterilizing pot influent hopper (25)).Opening centrifugal barrel electric motor carries out centrifugal, is thrown away by DEPC water in rifle head and EP pipe.Regulate high-pressure sterilizing pot air pressure and time, autoclaving twice, again open centrifugal barrel electric motor and carry out centrifugal, the water of condensation produced in rifle head during sterilizing and EP pipe is thrown away.DEPC water cycle uses repeatedly, is pumped into by DEPC water after heating in high-pressure sterilizing pot makes DEPC decompose and opens water discharge valve discharge.
Claims (7)
1. possess nuclease deactivation instrument and the application thereof of diethylpyrocarbonate isolation features, it is characterized in that the structure of described RNase deactivation instrument comprises: DEPC dilution and conservation tank (1), high-pressure sterilizing pot (2), base (3), universal wheel (4), connection valve (5), electric motor (6), air pump (7), DEPC well (8), conservation tank water intaking valve (9), conservation tank influent hopper (10), paddle (11), rifle head carriage (12), water discharge valve (13), vent valve (14), Autoclave top cover (15), heating tube (16), centrifugal barrel (17), rifle head carriage snap close (18), EP pipe fixed bin baffle plate (20), EP pipe fixed bin (21), rifle first gear plate (22), rifle head box panel (23), high-pressure sterilizing pot influent hopper (25), conservation tank and high-pressure sterilizing pot are arranged on the base of band universal wheel, air pump and conservation tank water intaking valve are arranged on above conservation tank, and connection valve is arranged on the below of conservation tank and high-pressure sterilizing pot, and control DEPC water flows between conservation tank and high-pressure sterilizing pot, centrifugal barrel is arranged on high-pressure sterilizing pot inside, and bottom connects electric motor, dismountable rifle head carriage is installed in centrifugal barrel, in centrifugal barrel, below is provided with EP pipe fixed bin (21).
2. the nuclease deactivation instrument possessing diethylpyrocarbonate isolation features according to claim 1, is characterized in that this RNase deactivation instrument can carry out RNase deactivation to batch to rifle head and EP pipe.
3. the nuclease deactivation instrument possessing diethylpyrocarbonate isolation features according to claim 1, it is characterized in that air pump can realize respectively conservation tank pressurization and air-extraction function under electric motor rotates and reverse drive, thus realize DEPC water and flow between conservation tank and high-pressure sterilizing pot.
4. according to claim 1 a kind of efficient and possess the RNase deactivation instrument of DEPC isolation features, it is characterized in that water pump (24) is arranged on conservation tank and autoclaving pot bottom pipe connecting, water pump can substitute air pump, has the function that control DEPC water flows between conservation tank and high-pressure sterilizing pot.
5. the nuclease deactivation instrument possessing diethylpyrocarbonate isolation features according to claim 1, is characterized in that being provided with dismountable rifle head carriage becoming 70-85 degree inclination angle with centrifugal barrel in centrifugal barrel.
6. rifle head carriage according to claim 5, is characterized in that rifle head carriage shape is as fallen " recessed " font, has aperture above rifle head carriage.
7. the nuclease deactivation instrument possessing diethylpyrocarbonate isolation features according to claim 1, is characterized in that EP pipe fixed bin is arranged to the large spaces such as 4 or 8.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510684453.5A CN105273976B (en) | 2015-10-22 | 2015-10-22 | Possesses the nucleic acid enzyme-deactivating instrument of pyrocarbonic acid diethyl ester isolation features |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510684453.5A CN105273976B (en) | 2015-10-22 | 2015-10-22 | Possesses the nucleic acid enzyme-deactivating instrument of pyrocarbonic acid diethyl ester isolation features |
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| Publication Number | Publication Date |
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| CN105273976A true CN105273976A (en) | 2016-01-27 |
| CN105273976B CN105273976B (en) | 2017-08-04 |
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| CN201510684453.5A Active CN105273976B (en) | 2015-10-22 | 2015-10-22 | Possesses the nucleic acid enzyme-deactivating instrument of pyrocarbonic acid diethyl ester isolation features |
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| CN205152201U (en) * | 2015-10-22 | 2016-04-13 | 甘肃农业大学 | High -efficient and possess DEPC isolating function's RNase deactivation appearance |
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2015
- 2015-10-22 CN CN201510684453.5A patent/CN105273976B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2149900Y (en) * | 1993-01-14 | 1993-12-22 | 张本文 | Device for boiling sucker |
| CN201361282Y (en) * | 2009-03-17 | 2009-12-16 | 邢庆华 | Medical disinfection container for easily floating articles |
| CN201510542U (en) * | 2009-10-14 | 2010-06-23 | 中国人民解放军第四军医大学 | Reverse transcription-polymerase chain reaction article sterilizing box |
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| CN105273976B (en) | 2017-08-04 |
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Inventor after: Tang Xun Inventor after: Xu Pingping Inventor after: Zou Duojiao Inventor after: Zhang Dongmin Inventor after: Chen Hanfang Inventor after: Duan Niane Inventor after: Xiao Wei Inventor after: Peng Xuelian Inventor after: Yin Guixiang Inventor after: Zhang Jianying Inventor after: Peng Liling Inventor after: Si Huaijun Inventor after: Liu Buming Inventor after: Hu Huamei Inventor after: He Yuetao Inventor after: Zeng Youling Inventor after: Zhang Weilong Inventor after: Liang Zaomei Inventor after: Liang Jiaoli Inventor after: Ruan Qingzhen Inventor after: Zhang Guodong Inventor after: Yang Jiangwei Inventor after: Zhang Ning Inventor after: Li Shigui Inventor after: Zhang Li Inventor after: Zhao Shirong Inventor after: Lei Yu Inventor before: Tang Xun Inventor before: Xu Pingping Inventor before: Si Huaijun Inventor before: Zhang Guodong Inventor before: Yang Jiangwei Inventor before: Zhang Ning Inventor before: Li Shigui Inventor before: Zhang Li Inventor before: Zhao Shirong Inventor before: Lei Yu |
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Effective date of registration: 20171121 Address after: 6 417A room, building No. 1188, west two ring road, Shengze Town, Wujiang District, Jiangsu, Suzhou, 215228 Patentee after: Suzhou hair textile Co., Ltd. Address before: 730070 Anning District, Gansu, Lanzhou Camp Village Gate No. 1 Patentee before: Gansu Agricultural University |
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Effective date of registration: 20181012 Address after: 215200 No. 1188 west two ring road, Shengze Town, Wujiang District, Suzhou, Jiangsu. Patentee after: Suzhou Shengze science and Technology Pioneer Park Development Co., Ltd. Address before: 215228 417A, 6 building, 1188 west two ring road, Shengze Town, Wujiang District, Suzhou, Jiangsu. Patentee before: Suzhou hair textile Co., Ltd. |