CN109609500A

CN109609500A - Utilize the method for the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle

Info

Publication number: CN109609500A
Application number: CN201910102619.6A
Authority: CN
Inventors: 赵仲玖; 廖东升; 邱坤; 赵伟
Original assignee: Chengdu Doosheng Biotechnology Co Ltd
Current assignee: Chengdu Doosheng Biotechnology Co Ltd
Priority date: 2019-02-01
Filing date: 2019-02-01
Publication date: 2019-04-12
Anticipated expiration: 2039-02-01
Also published as: WO2020156048A1; CN109609500B

Abstract

The method that the present invention discloses animal vegetable tissue ablatograph rapidly extracting RNA of the utilization based on ultrasonic principle, 1) first with the tissue or cell of extracting solution A preservation；Extracting solution B is added in the tissue or cell saved again to extracting solution A, mixes and obtains mixture；2) when extracting RNA from tissue, mixture is subjected to ultrasonication；3) from cell extract RNA when, directly carry out following step 5) in operation；4) mixture after ultrasonication is mixed by inversion；5) chloroform centrifugation is added, takes supernatant liquid from the mixture of cell extraction RNA or the mixture of step 4) in step 3)；6) isopropanol is added in supernatant liquid and is centrifuged acquisition sediment again；7) ethanol solution is added into sediment, is centrifuged after mixing, discards liquid；8) it is eventually adding water dissolution.The present invention uses ablation of tissue instrument ablation tissue, and rate is fast, avoid historrhexis not exclusively, the interference of endogenous RNA enzyme, and shorten extraction time, improve the extraction efficiency of RNA.

Description

Utilize the method for the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle

Technical field

The invention belongs to biologic product technology fields, and in particular to utilize the animal vegetable tissue ablatograph based on ultrasonic principle The method of rapidly extracting RNA.

Background technique

Separation and purifying RNA from tissue and cell are often required in modern molecular biology experiment, clinical molecular diagnosis, and The quality height of RNA usually will affect the success or failure of the molecular biology experiments such as the library cDNA, RT-PCR and Northern Blot.

Existing frequently-used method is to first pass through organization protection's liquid to protect tissue, then using extracting solution to tissue or thin RNA in born of the same parents is extracted.Common protected mode includes protection of liquid nitrogen and protection liquid protection, and the common protection liquid includes RNA later reagent and RNA safety reagent.

Wherein, not only required condition is harsh for protection of liquid nitrogen mode, and higher cost, is suitable only for provisional organization protection. And liquid is protected usually to be also required to be stored at low temperature, and wherein RNA later solution and TRIzol, silicagel column and paramagnetic particle method Chemical component it is incompatible, in RNA later solution contain isothiocyanic acid ammonium, when tissue block takes out from RNA later solution It is different in RNA later solution if not cleaning up the RNA later solution having on tissue block when for extracting RNA Ammonium thiocyanate can in extracting solution phenol, in conjunction with SDS or reaction generates precipitating, cause extracting solution ingredient and concentration to change, cracking It is not thorough, reduces the purity and yield of RNA.Therefore, usual before extraction using the tissue or cell of RNA later solution preservation It needs repeatedly to be cleaned, waste time and manpower, reduce extraction efficiency.

And RNA is extracted then usually using TRIzol method, silicagel column and paramagnetic particle method etc., but the generally existing RNA of these methods is received The problem that rate is low, impurity is seriously polluted is not able to satisfy the experiment such as subsequent gene diagnosis, analyzing biochips, gene expression analysis Quality requirement.

In addition, the shattering process of tissue block also strong influence the quality of RNA.The breaking method of common tissue block includes Liquid nitrogen grinding method, homogenate method (manual homogenization, machine homogenate, ultrasound homogenate) and multigelation method.The group that liquid nitrogen grinding method needs It is larger to knit block, and more difficult thoroughly smashes tissue block；The more difficult thorough inhibition endogenous RNA enzyme activity of homogenate method, be easy to cause interior The degradation of source property；The shortcomings that multigelation method is also to be difficult to thoroughly crack bulk tissue, and endogenous RNA enzyme activity is difficult to inhibit. Therefore, to the RNA extraction of animal derived tissue block, there are complicated for operation, broken incomplete, endogenous RNA activity to be difficult to inhibit Difficulty causes the extraction quality of RNA and yield to be difficult to ensure.

Summary of the invention

In order to solve the above problems existing in the present technology, it is an object of that present invention to provide utilize moving based on ultrasonic principle The method of plant tissue ablatograph rapidly extracting RNA.

The technical scheme adopted by the invention is as follows:

Using the method for the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle, comprising steps of

G1. tissue or cell first are saved with extracting solution A, the extracting solution A include 1.2-2.0mol/L guanidine thiocyanate and The guanidine hydrochloride of 0.5-1.5mol/L, and pH is 4；

G2. extracting solution B is added in the tissue or cell saved again to extracting solution A, is mixed by inversion, obtains mixture, it is described Extracting solution B includes with the water saturation phenol of the 30-65vt% of volume fraction and the glycerol of 9-15vt%, and surplus is solvent；

Extracting solution B further includes with the thiocyanic acid of the guanidine thiocyanate of the 0.20-1mol/L of molar concentration meter, 0.4-0.8mol/L The surfactant of ammonium and 0.0073-0.018mol/L, and pH is adjusted to 4；

When G3. extracting RNA from tissue, the mixture that step G2 is obtained utilizes the animal vegetable tissue based on ultrasonic principle Ablatograph carries out ultrasonication；Operation when extracting RNA from cell, in directly progress following step G5；

G4. the mixture after ultrasonication is mixed by inversion；

G5. chloroform is added from the mixture of the mixture of cell extraction RNA or step G4 in step G3, after mixing from The heart takes supernatant liquid；

G6. isopropanol is added in supernatant liquid, is centrifuged after mixing, discards liquid, obtains sediment；

G7. ethanol solution is added into sediment, is centrifuged after mixing, discards liquid；

G8. water dissolution is added after ethyl alcohol volatilization；

The animal vegetable tissue ablatograph based on ultrasonic principle, comprising: power supply, DCDC converting unit, main control unit, DCDC power adjustment unit, driving unit, voltage transformation unit, resonant element, ultrasonic transducer and sampling unit；

The power supply gives animal vegetable tissue ablation after converting voltage by DCDC converting unit and DCDC power adjustment unit Instrument uses；Main control unit, output the first pwm signal control DCDC power adjustment unit exports adjustable voltage, and is become by acquisition The feedback of the output end loop voltage of unit and the sampling unit of electric current is pressed, the second pwm signal of two-way complementary duty cycle is exported To driving unit and pass through voltage transformation unit transformation, is then adjusted by the resonant element being set in the output end loop of voltage transformation unit Resonance point to ultrasonic vibrator resonance point so that be set to voltage transformation unit output end loop in ultrasonic vibrator work in resonance shape State.

Ultrasonic vibrator includes ultrasonic transducer and passes after the vibration of ultrasonic transducer input is changed amplitude Amplitude transformer.

Based on the above technical solution, the surfactant in the extracting solution B is lauryl sodium sulfate.

Based on the above technical solution, the extracting solution A uses salt acid for adjusting pH value, and the extracting solution B uses second Acid for adjusting pH value.

Based on the above technical solution, further include in the extracting solution B molar concentration be 6.4mmol/L- 19.20mmol/L beta -mercaptoethanol.

Based on the above technical solution, further include in the extracting solution B molar concentration be 0.05-0.15mmol/L Potassium citrate.

Based on the above technical solution, further include in the extracting solution B molar concentration be 74.62 μm of ol/L- 223.87 the bromophenol blue of μm ol/L.

Based on the above technical solution, further include in the extracting solution B molar concentration be 0.01-2mol/L acetic acid Sodium.

Based on the above technical solution, further include in the extracting solution B volume fraction be 5-25% isopropanol.

Based on the above technical solution, further include in the extracting solution B molar concentration be 0.689mmol/L- 68.9mmol/L 8-hydroxyquinoline.

Based on the above technical solution, further include in the extracting solution B molar concentration be 0.5-1000mmol/L DTT。

Based on the above technical solution, further include in the extracting solution B molar concentration be 0.5-5mol/L urea.

Based on the above technical solution, further include in the extracting solution B molar concentration be 0.1-500mmol/L EDTA。

Based on the above technical solution, further include in the extracting solution B molar concentration be 0.01-10mmol/L second Sour ammonium.

Based on the above technical solution, the water in the step G8 is distilled water, deionized water or ultrapure water.

Based on the above technical solution, the gross mass of the extracting solution A and extracting solution B is tissue or cell quality 100-150 times.

Based on the above technical solution, in the step G3, ultrasonic power 0.5-1000W, ultrasonic time is 0.5-30s。

Based on the above technical solution, in the step G5, the volume of chloroform is the 0.1-0.4 of volume of mixture Times.

Based on the above technical solution, in the step G6, the volume ratio of isopropanol and supernatant liquid is 1:1.

Firstly, extracting solution main component of the invention is guanidine thiocyanate and guanidine hydrochloride, the guanidine thiocyanate is a kind of organic Compound, molecular formula C₂H₆N₄S is mainly used for biological medicine, chemical reagent etc..

Guanidine thiocyanate is to solve even agent, strength protein denaturant, destroy secondary protein structure simultaneously；Cell is mainly played to split Solution effect promotes nucleoprotein and nucleic acid separation；And enzyme inhibitor effect is played, have and saves tissue effect.

And the guanidine hydrochloride is a kind of block white or yellowish, as the strong denaturation extracted in cell total rna Agent.Guanidine hydrochloride solution can dissolve protein, and eucaryotic cell structure is caused to be destroyed, and nucleoprotein secondary structure is destroyed, under dissociating from nucleic acid Come, in addition, RNA enzyme can be inactivated by reducing agents such as guanidine hydrochlorides, it, can be from rich in RNase tissue as the inhibitor of nuclease Extract RNA.That is, guanidine thiocyanate can crack tissue or cell, but RNA is able to suppress by increased guanidine hydrochloride simultaneously The activity of enzyme, to achieve the effect that cracking and save.

It is stored at room temperature by the quantitative configuration of guanidine thiocyanate and guanidine hydrochloride to provide certain time to tissue or cell Effect can generally be saved more than 7 days at normal temperature.

Sodium dihydrogen phosphate, sodium chloride and magnesium chloride can be used as buffer system in extracting solution, to keep salt ion Concentration.And water-saturated phenol wherein included has the function of lytic cell and protein precipitation, and removal DNA is helped when in acidity； And ammonium thiocyanate is colourless crystallization, easily deliquesces, soluble easily in water and ethyl alcohol, is dissolved in methanol and acetone, is practically insoluble in chloroform and second Acetoacetic ester, can precipitating proteins, thus achieve the effect that separate impurity；The surfactant use SDS, i.e., ten two Sodium alkyl sulfate, having makes protein-denatured effect；And guarantee extraction quality is mainly played to 3.5-4.5 by adjusting pH value Effect will affect yield because of the too low purity that can mainly reduce extraction of pH, and excessively high.

Increased beta -mercaptoethanol, it has both the functional group of ethylene glycol and dithioglycol, is volatile liquid, has relatively strong Strong penetrating odor.β ME is commonly used in the reduction of disulfide bond, can be used as the antioxidant in biological experiment.It is wide The reason of general use is that its hydroxyl enables it to be dissolved in the water, and reduces its volatility.It is main in the present invention to rise To the disulfide bond destroyed in RNase, to play inhibitory effect.

Increased potassium citrate is a kind of common buffer, it is a kind of white, slightly hygroscopic crystalline powder.Nothing It is smelly, there is the taste of physiological saline, it is soluble easily in water, it is slow to be dissolved in glycerol, alcohol is not dissolved in, taste is salty and cool.Make stabilizer and pH buffer Deng, participation buffer system, the inhibitor as enzyme.

The molecular formula of increased bromophenol blue is C₁₉H₁₀Br₄O₅S arrives brownish-yellow powder to be light yellow, refers to usually as electrophoresis Show dyestuff, and is similarly used as color indicator in formula of the invention.In centrifugal process, bromophenol blue is with DNA and albumen Matter is sunk, and may be used to indicate whether RNA isolation is completed.

Increased sodium acetate can carbohydrate in RNA extraction process in precipitation solution, to reach the effect for removing impurity Fruit increases the purity of the RNA of extraction.

Increased isopropanol is a kind of organic compound, is colourless transparent liquid, makes RNA by the hydrophobic effect of-OH Hydrophilic radical in chain is protected, while having the function of restoring upper liquid RNA.

Increased 8-hydroxyquinoline can equally inhibit RNase, and inhibiting effect can be enhanced by being used in combination with chloroform.

Increased DTT is a kind of reducing agent, consistent with the effect of mercaptoethanol, primarily serves two sulphur destroyed in RNase Key, to play inhibitory effect.

Increased urea is a kind of organic compound of white crystalline；High concentration urea can be protein denaturation, and It can equally inhibit RNase active.

Increased EDTA is a kind of chelate divalent metal object, equally the inhibitor as enzyme.

Increased ammonium acetate skeleton symbol is CH₃COONH₄, also known as ammonium acetate.It is a kind of white triangles for having acetic acid odor Crystal can be used as analytical reagent and preservative for meat.In the present invention precipitating RNA extract in the protein that encounters, but ammonium root from Son is easy to form white precipitate with thiosulfate ion.

Wherein, extracting solution A can be 24 hours with room temperature preservation tissue, guarantee that RNA is non-degradable, that is, do not have to cryo-conservation, are detached from Cold chain.

The present invention uses ultrasonication tissue, and rate is fast, and it is incomplete, endogenous RNA enzyme to not only avoid historrhexis Interference, and extraction time is greatly shortened, further improve the extraction efficiency of RNA.

It is worth noting that therefore remaining liq component is with volume wherein what is calculated with molar concentration is powder ingredients Score calculate, and powdered ingredients because be dissolved in solvent thus its volume change can be neglected.

It is described to be based on ultrasonic principle using the method for the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle Animal vegetable tissue ablatograph DCDC power adjustment unit include: switch control signal receiving end connection main control unit reception open The switch control signal for closing control signal receives circuit；

First pwm signal receiving end connects the first pwm signal reception circuit that main control unit receives the first pwm signal；

Power supply is converted into adjustable voltage output by the switch control signal and the first pwm signal of main control unit transmission 4th DCDC converting unit.

Preferably, the 4th DCDC converting unit of the animal vegetable tissue ablatograph includes: successively to handle supply voltage The 4th voltage input filtering, the 4th voltage conversion chip U1 and the 4th voltage output filtering, the 4th voltage input filtering includes It is connected to the filter capacitor of power end, the upper tube driving signal reference point pin SW of the 4th voltage conversion chip U1 passes through energy storage electricity Sense L1 is connected to adjustable voltage output end, and divider resistance R32 and R47 the progress voltage sample for being connected on adjustable voltage output end are defeated Enter the reference voltage pin FB to the 4th voltage conversion chip U1, the upper tube driving signal reference point of the 4th voltage conversion chip U1 Pin SW is also connected with sustained diode 2, and enable pin EN is also connected with resistance R33, resistor timing/external clock pin RT/CLK is also connected with frequency divider resistance R85, and frequency compensation pin COMP is also connected with defeated for adjusting loop, burning voltage Capacitor C51, C49 and resistance R46 out, wherein capacitor C49 is in parallel with concatenated capacitor C51 and resistance R46, and the 4th voltage is defeated Filtering includes the filter capacitor for being connected to adjustable voltage output end out；

It includes: the electricity being series between converted voltage and switch control signal receiving end that switch control signal, which receives circuit, R104 and R105 is hindered, emitter connects the node between converted voltage, base stage connection resistance R104 and R105 and collector connects The triode Q12 for meeting the enable pin EN of the 4th voltage conversion chip U1, when switch control signal is L, triode Q12 is led Logical, the 4th voltage conversion chip U1 opens work；

It includes: RC filtering and the first voltage follower for successively handling the first pwm signal that first pwm signal, which receives circuit, U3B, received first pwm signal in the first pwm signal receiving end are input to first voltage after being filtered by RC filtering and follow The non-inverting input terminal of device U3B, the output end of first voltage follower U3B by divider resistance R54 be connected to divider resistance R32 and Node between R47.

Preferably, the animal vegetable tissue ablatograph further includes the adjustable voltage for acquiring DCDC power adjustment unit Output voltage sampling unit, the adjustable voltage of acquisition is sent to main control unit by second voltage follower U3A.

Preferably, the driving unit of the animal vegetable tissue ablatograph includes that the first driving unit and the second driving are single Member, the second pwm signal receiving end N of the first driving unit receive the second pwm signal of the first via that main control unit is sent, and first drives The Same Name of Ends of the primary coil of the output end connection voltage transformation unit of moving cell, the second pwm signal receiving end P of the second driving unit Receive second the second pwm signal of tunnel that main control unit is sent, the primary side line of the output end connection voltage transformation unit of the second driving unit The different name end of circle is led by received the first driving of the second pwm signal of the first via control metal-oxide-semiconductor Q6 of the second pwm signal receiving end N Logical/cut-off, by received the second pwm signal of second tunnel control the second driving metal-oxide-semiconductor Q2 cut-off of the second pwm signal receiving end P/ Conducting.

Preferably, the voltage transformation unit of the animal vegetable tissue ablatograph is push-pull transformer, DCDC power adjustment unit Adjustable voltage output end connection push-pull transformer primary side first coil different name end and the second coil Same Name of Ends；

First driving unit includes the resistance R10 and R14 being series between the second pwm signal receiving end N, grid Node, source electrode between pole connection resistance R10 and R14 are grounded and drain electrode is the output end of the first driving unit and change is recommended in connection First driving metal-oxide-semiconductor Q6 of the Same Name of Ends of the primary side first coil of depressor；

Second driving unit includes the resistance R5 and R13 being series between the second pwm signal receiving end P, grid Node, source electrode between pole connection resistance R5 and R13 are grounded and drain electrode is the output end of the second driving unit and change is recommended in connection The second driving metal-oxide-semiconductor Q2 at the different name end of the second coil of primary side of depressor.

When received the second pwm signal of the first via of the second pwm signal receiving end N is H and the second pwm signal receiving end P connects When second the second pwm signal of tunnel received is L, control the first driving metal-oxide-semiconductor Q6 conducting, the second driving metal-oxide-semiconductor Q2 cut-off, primary side the The input end loop of one coil is connected and input current direction is the different name end of primary side first coil to Same Name of Ends, primary side second The input terminal loop cutoff of coil, the output current direction of the output end loop of secondary coil are the Same Name of Ends of secondary coil to different Name end.

When received the second pwm signal of the first via of the second pwm signal receiving end N is L and the second pwm signal receiving end P connects When second the second pwm signal of tunnel received is H, control the first driving metal-oxide-semiconductor Q6 cut-off, the second driving metal-oxide-semiconductor Q2 conducting, primary side the The input end loop of two coil is connected and input current direction is the Same Name of Ends of the second coil of primary side to different name end, primary side first The input terminal loop cutoff of coil, the output current direction of the output end loop of secondary coil are the different name end of secondary coil to same The DC voltage of input, is converted into up to a hundred volts of frequency 30KHz, voltage of AC wave shape by name end, is mentioned for rear class resonant element For condition.

Preferably, the resonant element of the animal vegetable tissue ablatograph is LC series resonance, LC series resonance includes string The inductance T1 and capacitor C1 being coupled in the output end loop of voltage transformation unit.

Preferably, the ultrasonic transducer of the animal vegetable tissue ablatograph is set to the output of voltage transformation unit by interface J2 In end loop.

Preferably, the sampling unit of the animal vegetable tissue ablatograph includes the output end loop electricity for acquiring voltage transformation unit The current sampling unit of the output end loop current of the voltage sampling unit and acquisition voltage transformation unit of pressure, correspondingly, voltage transformation unit Output end loop be equipped with multiple concatenated sampling resistors；By the voltage sampling port for being connected to the intermediate node of multiple sampling resistors Collection voltages signal is then forwarded to voltage sampling unit and is filtered and amplifies, and is then forwarded to main control unit and does resonance tune Section；By be connected on voltage transformation unit output end loop current sampling port acquire current signal be sent to current sampling unit into Row filtering and amplification, are then forwarded to main control unit and do resonance adjusting.

Preferably, the main control unit of the animal vegetable tissue ablatograph includes main control chip U2 and main control chip periphery electricity Road, main control chip U2 exports the first pwm signal control DCDC power adjustment unit and exports adjustable voltage, and passes through acquisition transformation list The feedback of the sampling unit of the output end loop voltage and electric current of member, the second pwm signal for exporting two-way complementary duty cycle extremely drive Moving cell.

Preferably, the main control unit further includes companion chip U1 and companion chip peripheral circuit, the master of main control unit Chip U2 is controlled by the feedback of the output end loop voltage of acquisition voltage transformation unit and the sampling unit of electric current, is sent out to companion chip U1 Send instruction, companion chip U1 receives the instruction of main control chip U2, exports the second pwm signal of two-way complementary duty cycle to driving list Member, meanwhile, the second pwm signal of two-way complementary duty cycle also feeds back to main control chip U2.

Preferably, the second pwm signal of the two-way complementary duty cycle also passes through the first signal amplification output electricity respectively Road and second signal amplification output circuit are sent to driving unit after amplifying.

Preferably, the animal vegetable tissue ablatograph further includes the display unit connecting with main control chip U2, display is single Member includes display chip J8 and the LCD display connecting with display chip J8.

Preferably, the animal vegetable tissue ablatograph further includes the key connecting with main control chip U2.

Preferably, the animal vegetable tissue ablatograph further includes the memory connecting with main control chip U2.

Preferably, the animal vegetable tissue ablatograph further includes the usb interface unit connecting with main control chip U2, USB Interface unit includes the USB chip ESD1 connecting with main control chip U2 and the interface J7 that connect with USB chip ESD1.

Preferably, the animal vegetable tissue ablatograph further includes the touch unit connecting with main control chip U2.

The invention has the benefit that

(1) extracting solution A of the invention may replace traditional RNA later solution, for saving tissue or cell, therefore, when When extracting RNA using extracting solution A and extracting solution B, does not need to clean tissue or cell, eliminate using RNA later Cleaning operation when solution saves, simplifies operating procedure, significantly improves extraction efficiency；

(2) each component proportion design in extracting solution of the present invention is reasonable, acts synergistically between each other, so that the RNA extracted Purity is high, high income, and protein contamination is low；

(3) animal vegetable tissue ablatograph of the invention, can using the control DCDC power adjustment unit output of the first pwm signal Voltage is adjusted, keeps power adjustable in 0.5~1000W, utilizes the output end loop voltage of acquisition voltage transformation unit and the sampling list of electric current The feedback of member exports the second pwm signal of two-way complementary duty cycle to driving unit, makes resonant ultrasonic frequency 20-200KHz It is adjustable, resonance point is adjusted to the resonance point of ultrasonic vibrator, so that ultrasonic vibrator work in resonant state, exports, energy is maximum, shakes Width is most strong, and using ultrasonication tissue, rate is fast, not only avoid historrhexis not exclusively, the interference of endogenous RNA enzyme, and And extraction time is greatly shortened, further improve the extraction efficiency of RNA；

(4) extracting solution A of the present invention, extracting solution B property are more stable, do not influence vulnerable to conditions such as temperature and humidity, and can be sufficiently With the use of, it can be achieved that laboratory is quick, the purpose of animal RNA is extracted in cheap, high quality, scale；

(5) the newly-increased guanidine hydrochloride of the present invention, has nucleic acid inhibitor effect, can effectively extract from rich in RNase tissue RNA, newly-increased sodium acetate have the carbohydrate effect encountered in precipitating RNA extraction, guarantee the RNA purity extracted, increase isopropanol newly: is logical The hydrophobic effect for crossing-OH protects the hydrophilic radical in RNA chain, while having the function of restoring upper liquid RNA；

(6) present invention has adjusted A liquid and saves tissue amount of reagent, and tissue can be used for room temperature preservation to 10 days or more, but It is directed to the liver organization rich in RNase, the RNA result extracted is normal, and yield is improved；In addition mainly exist RNase pollution aspect increases multiple groups ingredient, and RNA is prevented not to be degraded.

Detailed description of the invention

Fig. 1 is the present invention-embodiment system block diagram.

Fig. 2 is that the circuit of the present invention-embodiment control panel power supply, DCDC converting unit and DCDC power adjustment unit is former Reason figure.

Fig. 3 is the circuit diagram of the present invention-embodiment control panel main control unit.

Fig. 4 is the circuit diagram of the present invention-embodiment control panel sampling unit, display unit and companion chip.

Fig. 5 is the circuit diagram of the present invention-embodiment control panel interface.

Fig. 6 is the circuit diagram of the present invention-embodiment driving plate driving unit, voltage transformation unit and resonant element.

Fig. 7 is the agarose gel electrophoresis figure piece in the embodiment of the present invention 5.

Specific embodiment

With reference to the accompanying drawing and specific embodiment the present invention is further elaborated.

Embodiment 1:

As shown in figures 1 to 6, the animal vegetable tissue ablatograph based on ultrasonic principle of the present embodiment, including power supply, DCDC turn Change unit, main control unit, DCDC power adjustment unit, driving unit, voltage transformation unit, resonant element, ultrasonic transducer and sampling Unit.

Power supply, by making after DCDC converting unit and DCDC power adjustment unit conversion voltage to animal vegetable tissue ablatograph With；Main control unit, output the first pwm signal control DCDC power adjustment unit exports adjustable voltage, and passes through acquisition transformation list The feedback of the sampling unit of the output end loop voltage and electric current of member, the second pwm signal for exporting two-way complementary duty cycle extremely drive Moving cell simultaneously passes through voltage transformation unit transformation, then adjusts resonance by the resonant element being set in the output end loop of voltage transformation unit Point arrives the resonance point of ultrasonic vibrator, so that the ultrasonic vibrator being set in the output end loop of voltage transformation unit works in resonant state.

Animal vegetable tissue ablatograph is further elaborated.

Power supply is DC24V power supply, and DCDC converting unit includes the first DCDC converting unit, the 2nd DCDC converting unit and the Three DCDC converting units.

Wherein, the first DCDC converting unit include TVS protection, first voltage input filter, first voltage conversion chip and First voltage output filtering, DC24V power supply pass sequentially through TVS protection, first voltage input filter, first voltage conversion chip and First voltage output filtering is converted to 12V output voltage, supplies main control unit and driving unit uses.

Elaborate the circuit theory of the first DCDC converting unit: DC24V power supply successively pass through protection diode TVS1 into Row TVS protection, capacitor C59, C60 and the C57 for being connected in parallel on DC24V power end by three carry out voltage input filtering, then pass through Cross the enabled of divider resistance R48 and R51 progress voltage sample input the first voltage conversion chip U6 for being connected on DC24V power end The upper tube driving signal of pin EN, the model TPS54340 of first voltage conversion chip U6, first voltage conversion chip U6 are joined Examination point pin SW is connected to 12V output voltage terminal by energy storage inductor L2, is connected on the divider resistance R88 of 12V output voltage terminal The reference voltage pin FB that voltage sample is input to first voltage conversion chip U6 is carried out with R89, by being connected to 12V output electricity The capacitor C72 of pressure side filters voltage output, and the upper tube driving signal reference point pin SW of first voltage conversion chip U6 also connects It is connected to sustained diode 4, enable pin EN is also connected with soft start capacitor C79, resistor timing/external clock pin RT/ CLK is also connected with frequency divider resistance R50, and frequency compensation pin COMP is also connected with for adjusting loop, burning voltage output Capacitor C61, C62 and resistance R52, wherein capacitor C62 is in parallel with concatenated capacitor C61 and resistance R52.

2nd DCDC converting unit includes second voltage input filter, second voltage conversion chip and second voltage output filter Wave, 12V input voltage pass sequentially through second voltage input filter, second voltage conversion chip and second voltage output filtering conversion For 1.8V output voltage, supplies main control unit and use.

Elaborate the circuit theory of the 2nd DCDC converting unit: 12V input voltage successively passes through two, and to be connected in parallel on 12V defeated The capacitor C84 and C85 for entering voltage end carry out voltage input filtering, the model of second voltage conversion chip U9 The metal-oxide-semiconductor driving signal reference point pin SW of TLV62130ARGTR, second voltage conversion chip U9 are connected by energy storage inductor L3 To 1.8V output voltage terminal, the divider resistance R90 and R91 for being connected on 1.8V output voltage terminal carry out voltage sample and are input to second The reference voltage pin FB of voltage conversion chip U9, it is defeated to voltage by the capacitor C80 and C83 that are connected in parallel on 1.8V output voltage terminal It filters, the internal power power pin PVIN of second voltage conversion chip U9, internal control circuit power pin AVIN, enables out Pin EN is also connected with 12V Input voltage terminal, and soft start/tracking pin SS/TR is also connected with soft start capacitor C86, adjusts output Pin DEF and frequency configuration pin FSW are grounded, and output voltage acquisition pin VOS is connect with 1.8V output voltage terminal.

3rd DCDC converting unit includes tertiary voltage input filter, tertiary voltage conversion chip and tertiary voltage output filter Wave, 12V input voltage pass sequentially through tertiary voltage input filter, tertiary voltage conversion chip and tertiary voltage output filtering conversion For 3.3V output voltage, supplies main control unit and sampling unit uses.

Elaborate the circuit theory of the 3rd DCDC converting unit: 12V input voltage successively passes through two, and to be connected in parallel on 12V defeated The capacitor C89 and C90 for entering voltage end carry out voltage input filtering, the model of tertiary voltage conversion chip U10 The metal-oxide-semiconductor driving signal reference point pin SW of TLV62130ARGTR, tertiary voltage conversion chip U10 are connected by energy storage inductor L4 It is connected to 3.3V output voltage terminal, the divider resistance R92 and R94 for being connected on 3.3V output voltage terminal carry out voltage sample and be input to the The reference voltage pin FB of three voltage conversion chip U10, by being connected in parallel on the capacitor C87 and C88 of 3.3V output voltage terminal to electricity Pressure output filtering, the internal power power pin PVIN and internal control circuit power pin of tertiary voltage conversion chip U10 AVIN is also connected with 12V Input voltage terminal, and the divider resistance R93 and R102 by being connected on 12V Input voltage terminal carry out voltage and adopt Sample inputs the enable pin EN of tertiary voltage conversion chip U10, and the enable pin EN of tertiary voltage conversion chip U10 is also connected to The normal indication signal pin PG of the output of second voltage conversion chip U9, when the 2nd DCDC converting unit irregular working, the Three DCDC converting units then stop working, and soft start/tracking pin SS/TR is also connected with soft start capacitor C91, adjust efferent duct Foot DEF and frequency configuration pin FSW are grounded, and output voltage acquisition pin VOS is connect with 3.3V output voltage terminal.

DCDC power adjustment unit includes that switch control signal receives circuit, the first pwm signal receives circuit and the 4th DCDC converting unit, the 4th DCDC converting unit include the filtering of the 4th voltage input, the 4th voltage conversion chip and the 4th voltage Output filtering, the switch control signal and the first pwm signal that the 4th DCDC converting unit is sent by main control unit are electric by DC24V Source is converted to the output of 0-24V adjustable voltage.

Elaborate the circuit theory of DCDC power adjustment unit:

The circuit theory of 4th DCDC converting unit: DC24V power supply successively passes through three electricity for being connected in parallel on DC24V power end Hold C6, C7 and C41 and carries out voltage input filtering, the model TPS54340 of the 4th voltage conversion chip U1, the conversion of the 4th voltage The upper tube driving signal reference point pin SW of chip U1 is connected to adjustable voltage output end by energy storage inductor L1, is connected on adjustable The divider resistance R32 and R47 of voltage output end carry out the voltage reference tube that voltage sample is input to the 4th voltage conversion chip U1 Foot FB, capacitor C14, C40, C42, C54, C55 and C56 by being connected in parallel on adjustable voltage output end, which export adjustable voltage, to be filtered The upper tube driving signal reference point pin SW of wave, the 4th voltage conversion chip U1 is also connected with sustained diode 2, enable pin EN is also connected with resistance R33, and resistor timing/external clock pin RT/CLK is also connected with frequency divider resistance R85, and frequency is mended Repay pin COMP be also connected with for adjust loop, burning voltage output capacitor C51, C49 and resistance R46, wherein capacitor C49 is in parallel with concatenated capacitor C51 and resistance R46.

The circuit theory of switch control signal reception circuit: switch control signal receiving end connects main control unit and receives switch Signal is controlled, resistance R104 and R105 are series between 3.3V voltage and switch control signal receiving end, the transmitting of triode Q12 3.3V voltage, the node between base stage connection resistance R104 and R105 and the collector of pole connection connect the 4th voltage conversion chip The enable pin EN of U1, when switch control signal is L, triode Q12 conducting, the 4th voltage conversion chip U1 opens work.

The circuit theory of first pwm signal reception circuit: received first pwm signal in the first pwm signal receiving end passes through RC filtering is input to the non-inverting input terminal of first voltage follower U3B, the output end of first voltage follower U3B after being filtered The node between divider resistance R32 and R47 is connected to by divider resistance R54, wherein resistance R45, capacitor C63, resistance R40 With capacitor C52 composition RC filtering, the non-inverting input terminal of first voltage follower U3B, the first electricity are then input to by resistance R36 The non-inverting input terminal of pressure follower U3B is also connected with capacitor C50.

Animal vegetable tissue ablatograph further includes that the output voltage for acquiring the adjustable voltage of DCDC power adjustment unit is adopted The adjustable voltage of sample unit, acquisition is sent to main control unit by second voltage follower U3A.

It elaborates the circuit theory of output voltage sampling unit: being connected on the divider resistance R55 of adjustable voltage output end Voltage sample is carried out with R60 and the non-inverting input terminal of second voltage follower U3A is input to after being filtered by RC filtering, the The cathode voltage end of two voltage follower U3A connects 3.3V voltage, and the output end of second voltage follower U3A connects master control list Member, wherein resistance R59 and capacitor C39 composition RC filtering, 3.3V voltage are also connected with capacitor C44, second voltage follower U3A Output end be also connected with capacitor C43.

Driving unit includes the first driving unit and the second driving unit, wherein the first driving unit includes the first driving Metal-oxide-semiconductor Q6, the second driving unit include the second driving metal-oxide-semiconductor Q2.Second pwm signal receiving end N of the first driving unit is received The second pwm signal of the first via that main control unit is sent, the primary coil of the output end connection voltage transformation unit of the first driving unit Same Name of Ends, the second pwm signal receiving end P of the second driving unit receive second the second pwm signal of tunnel that main control unit is sent, the The different name end of the primary coil of the output end connection voltage transformation unit of two driving units, it is received by the second pwm signal receiving end N The second pwm signal of first via control the first driving metal-oxide-semiconductor Q6 conduction and cut-off, passes through the second pwm signal receiving end P received the Two the second pwm signals of tunnel control the second driving metal-oxide-semiconductor Q2 cut-off/conducting.

Elaborate the circuit theory of driving unit and voltage transformation unit: voltage transformation unit is push-pull transformer, DCDC power tune The different name end of the primary side first coil of the adjustable voltage output end connection push-pull transformer of unit and the Same Name of Ends of the second coil are saved, Resistance R10 and R14 be series between the second pwm signal receiving end N, first driving metal-oxide-semiconductor Q6 grid connect resistance R10 Node between R14, source electrode ground connection, drain for the first driving unit output end and connect the primary side first of push-pull transformer The Same Name of Ends of coil；Resistance R5 and R13 be series between the second pwm signal receiving end P, second driving metal-oxide-semiconductor Q2 grid Pole connect resistance R5 and R13 between node, source electrode ground connection, drain for the second driving unit output end and connection recommend transformation The different name end of the second coil of primary side of device.

Resonant element is set in the output end loop of push-pull transformer, and resonant element is LC series resonance, including is series at Inductance T1 and capacitor C1 in the output end loop of push-pull transformer.Inductance T1 uses EFD20.

Ultrasonic transducer is set in the output end loop of push-pull transformer, in the present embodiment, passes through interface J2 connection ultrasound Energy converter.

Sampling unit acquires the output end loop voltage and electric current of push-pull transformer, and sampling unit includes voltage sampling unit And current sampling unit.

Voltage sampling unit acquires the output end loop voltage of push-pull transformer, and the output end loop of push-pull transformer is equipped with Concatenated sampling resistor R1, R2, R3, R4 and R15, voltage sampling port are connected to the intermediate node of resistance R4 and R15, and by acquisition Voltage signal is sent to voltage sampling unit and is filtered, amplifies, and is then forwarded to main control unit and does resonance adjusting.

Elaborate the circuit theory of voltage sampling unit: the voltage signal of acquisition pass sequentially through RC filtering be filtered, The non-inverting input terminal of the first amplifier U7B, the inverting input terminal and output of the first amplifier U7B are input to after capacitor C67 progress blocking Negative feedback resistor R63 is connected between end then passes through resistance after the output end of the first amplifier U7B is filtered by RC filtering R72 is input to the non-inverting input terminal of first comparator U8A, be series at divider resistance R77 and R76 between 3.3V voltage and ground into Row voltage sample is input to the inverting input terminal of first comparator U8A, and the output end of first comparator U8A is connected by resistance R71 Main control unit is connect, the other end of the non-inverting input terminal of resistance R72 connection first comparator U8A also passes through resistance R70 connection master control Unit, wherein resistance R61 and capacitor C65 composition RC filtering is filtered the voltage signal of acquisition, resistance R62 and capacitor C66 Composition RC filtering is filtered the signal of the output end output of the first amplifier U7B, and the inverting input terminal of the first amplifier U7B is also gone here and there It is associated with resistance R64 and capacitor C70, the non-inverting input terminal of the first amplifier U7B is also connected with direct current 3.3V voltage, direct current 3.3V electricity Pressure generates the non-inverting input terminal that partial pressure is input to the first amplifier U7B after dividing by divider resistance R56, R58 and R57, partial pressure is also It is filtered by capacitor C64, wherein one end of resistance R58 connects the non-inverting input terminal of the first amplifier U7B and the other end connects resistance R56, resistance R56 connection 3.3V voltage, node and the other end between one end connection the resistance R56 and R58 of resistance R57 are grounded, The cathode voltage end of first comparator U8A connects 3.3V voltage, which passes through capacitor C76 and C77 filtering in parallel.

Current sampling unit acquires the output end loop current of push-pull transformer, and current sampling port is connected on push-pull transformer Output end loop, and the current signal of acquisition is sent to current sampling unit and is filtered, amplifies, be then forwarded to master control Unit does resonance adjusting.

Elaborate the circuit theory of current sampling unit: the current signal of acquisition pass sequentially through RC filtering be filtered, The non-inverting input terminal of the second amplifier U7A, the inverting input terminal and output of the second amplifier U7A are input to after capacitor C75 progress blocking Negative feedback resistor R73 is connected between end then passes through resistance after the output end of the second amplifier U7A is filtered by RC filtering R81 is input to the non-inverting input terminal of the second comparator U8B, be series at divider resistance R82 and R83 between 3.3V voltage and ground into Row voltage sample is input to the inverting input terminal of the second comparator U8B, and the output end of the second comparator U8B is connected by resistance R80 Main control unit is connect, the other end of the non-inverting input terminal of resistance R81 the second comparator of connection U8B also passes through resistance R79 connection master control Unit, wherein resistance R65 and capacitor C68 composition RC filtering is filtered the current signal of acquisition, resistance R69 and capacitor C69 Composition RC filtering is filtered the signal of the output end output of the second amplifier U7A, and the inverting input terminal of the second amplifier U7A is also gone here and there It is associated with resistance R74 and capacitor C78, the non-inverting input terminal of the second amplifier U7A is also connected with direct current 3.3V voltage, direct current 3.3V electricity Pressure generates the non-inverting input terminal that partial pressure is input to the second amplifier U7A after dividing by divider resistance R78, R66, R67 and R68, point Pressure is filtered by capacitor C71, wherein one end of resistance R68 connects the non-inverting input terminal of the second amplifier U7A and other end connection string It is coupled to the resistance R78 and R66 of 3.3V voltage end, node and the other end between one end connection the resistance R66 and R68 of resistance R67 Ground connection, by resistance R78 connection 3.3V voltage, which passes through electricity in parallel at the cathode voltage end of the second comparator U8B Hold C73 and C74 filtering.

Main control unit, output the first pwm signal control DCDC power adjustment unit exports adjustable voltage, and is become by acquisition The feedback of the output end loop voltage of unit and the sampling unit of electric current is pressed, the second pwm signal of two-way complementary duty cycle is exported The driving of metal-oxide-semiconductor Q6 and second metal-oxide-semiconductor Q2 is driven to the first of driving unit.

Main control unit includes main control chip U2 and main control chip peripheral circuit, wherein the model of main control chip U2 N32905U1DN, using ARM9 kernel, dominant frequency 200MHz, main control chip peripheral circuit includes system clock, reset etc..

N32905U1DNN3290x is based on ARM926EJ-S CPU core, is integrated with JPEG codec, cmos sensor Interface, 32 channel SPU (sound processing unit), ADC, DAC can meet various application demands, while save BOM cost. ARM926@200MHz, synchronous dram, 2D BitBLT accelerator, cmos image sensor interface, LCD panel interface. N32905U1DNN3290x maximum resolution is XVGA (1,024x768)@TFT LCD panel.2D BitBLT accelerator accelerates figure Shape calculates, and makes render smooth, CPU is unloaded, to save power consumption.

It is required to meet the different of total system BOM cost, various sizes of DRAM and the main SoC of N3290x are stacked into one A encapsulation, i.e. multi-chip package (MCP).N32905U1DNN3290x especially uses 1Mbitx16 3.3V SDRAM to design. N32905U1DNN3290x especially uses 4Mbitx16 1.8V DDR SDRAM to design.One 16Mbitx16 1.8V DDR2SDRAM is stacked on inside N32905U1DNN3290x, is designed with ensuring higher performance and reducing system to the maximum extent Work, such as EMI and noise coupling.By using the double-deck PCB and damping resistance is eliminated, EMI protection component etc. can reduce total BOM cost.

Above technical scheme sufficiently discloses and implementable, of the invention animal vegetable tissue ablatograph, utilizes the first PWM Signal controls DCDC power adjustment unit and exports adjustable voltage, keeps output power adjustable in 0.5~1000W, utilizes acquisition transformation The feedback of the sampling unit of the output end loop voltage and electric current of unit exports the second pwm signal of two-way complementary duty cycle extremely Driving unit keeps resonant ultrasonic frequency 20-200KHz adjustable, the resonance point of adjusting resonance point to ultrasonic vibrator, so that ultrasonic It is most strong to export energy maximum, amplitude in resonant state for oscillator work.

Embodiment 2:

On the basis of embodiment 1, the present embodiment further discloses the animal vegetable tissue ablatograph based on ultrasonic principle, In specific application, main control unit of the invention further includes companion chip U1 and companion chip peripheral circuit, companion chip U1's Model STM32F031G4U6.

STM32F031G4U6 is characterized in: kernel:32- M0 CPU, frequency are up to 48MHz；Storage: 16 to 32KB flash memory, the 4KB SRAM with hardware parity；CRC computing unit；Resetting and power management number and I/O Power supply: 2.0 to 3.6V, analog power: VDDA=is powered on/off reset (POR/PDR) from VDD to 3.6V, programmable voltage inspection Survey device (PVD), low-power consumption mode: sleep stops and supports, the VBAT power supply for RTC and back-up registers；Clock management: 4 To 32MHz crystal oscillator, for the 32kHz oscillator of the RTC with calibration, the inside 8MHz RC with x6PLL option is interior Portion's 40kHz RC oscillator；Most 39 quick I/O, all to map to external interrupt vector, most 26 I/O have 5V Tolerance；5 channel DMA controllers；1 × 12,1.0 μ sADC (most 10 channels), conversion range: 0 to 3.6V, by simulation electricity Source is separated to 3.6V from 2.4；Most 9 timers, the 7 advanced control timer in channel of the position 1x16 are exported for 6 channel PWM, tool There are dead time, power generation and emergent stopping, 1x32 and 1x16 bit timing device, most 4 IC/OC can be used for IR control decoding, 1x16 bit timing device, band 2 IC/OC, 1 OCN, dead time and emergent stopping, 1x16 bit timing device, band IC/OC and OCN, Dead time, emergent stopping and the modulator door for IR control, the 1x16 bit timing device with 1 IC/OC, independence and system WatchDog Timer, SysTick timer: 24 Down-counters；Calendar RTC has the function of alarm and periodic wakeup from stopping Only/standby；1 I2C interface of communication interface supports quick mode to add (1Mbit/s), and 20mA electric current absorbs, SMBus/PMBus, From Stop awakening mode, 1x USART supports main synchronization, SPI and modem control, ISO7816 interface, LIN, IrDA function Can, the detection of auto-baud rate and arousal function, 1x SPI (18Mbit/s), 4 to 16 programmable bit frames, band I2S interface；String Line debugs (SWD)；96 unique ID；Extended temperature range: -40 to+105 DEG C；All packagings2。

The main control chip U2 of main control unit passes through the output end loop voltage of acquisition voltage transformation unit and the sampling unit of electric current Feedback, to companion chip U1 send instruct, companion chip U1 receive main control chip U2 instruction, output two-way frequency be 30KHz, the first of the second pwm signal of complementary duty cycle to driving unit drive the driving metal-oxide-semiconductor Q2 of metal-oxide-semiconductor Q6 and second, together When, two-way frequency is 30KHz, and the second pwm signal of complementary duty cycle also feeds back to main control chip U2.Meanwhile first comparator The signal of the output end of the output end of U8A and the second comparator U8B output is sent to companion chip U1.

Two-way frequency is 30KHz, and the second pwm signal of complementary duty cycle also passes through the first signal amplification output circuit respectively The the first driving metal-oxide-semiconductor Q6 and the second driving MOS of driving unit are sent to after amplifying with second signal amplification output circuit Pipe Q2.

Elaborate the circuit theory of the first signal amplification output circuit: the second pwm signal passes through concatenated divider resistance The base stage that R49 and R53 inputs the first amplifying triode Q5 amplifies, and the collector of the first amplifying triode Q5 passes through resistance R34 connection 12V voltage, the emitter ground connection of the first amplifying triode Q5, the collector of the first amplifying triode Q5 and first defeated The base stage of triode Q2 and the second output triode Q4 connect out, and the first output triode Q2 is p-type triode, the second output three Pole pipe Q4 is N-type triode, and the collector of the first output triode Q2 connects 12V voltage, amplified the 2nd PWM of first via letter It number exports from the emitter connecting node of the emitter of the first output triode Q2 and the second output triode Q4 to first and to drive The second pwm signal receiving end N of dynamic metal-oxide-semiconductor Q6.

When the input of the second pwm signal is H, the first output triode Q2 conducting, the second output triode Q4 cut-off；When When the input of two pwm signals is L, thus the first output triode Q2 cut-off, the second output triode Q4 conducting exports amplified The second pwm signal of the square wave first via.

Elaborate the circuit theory of second signal amplification output circuit: the second pwm signal passes through concatenated divider resistance The base stage that R86 and R87 inputs the second amplifying triode Q8 amplifies, and the collector of the second amplifying triode Q8 passes through resistance R84 connection 12V voltage, the emitter ground connection of the second amplifying triode Q8, the collector and third of the second amplifying triode Q8 are defeated The base stage of triode Q6 and the 4th output triode Q7 connect out, and third output triode Q6 is p-type triode, the 4th output three Pole pipe Q7 is N-type triode, and the collector of third output triode Q6 connects 12V voltage, amplified the 2nd PWM of second tunnel letter It number exports from the emitter connecting node of the emitter of third output triode Q6 and the 4th output triode Q7 to second and to drive The second pwm signal receiving end P of dynamic metal-oxide-semiconductor Q2.

When the input of the second pwm signal is H, third output triode Q6 conducting, the 4th output triode Q7 cut-off；When When the input of two pwm signals is L, thus third output triode Q6 cut-off, the 4th output triode Q7 conducting exports amplified The second pwm signal of the second tunnel of square wave.

Main control chip U2 is also connected with display unit, and display unit includes display chip J8 and connect with display chip J8 LCD display, the model FPC050 of display chip J8, display chip J8 connection 3.3V voltage, the signal of main control chip U2 LCD_BL is sent to display chip J8 after amplifying by metal-oxide-semiconductor Q1.

Main control chip U2 is also connected with key, and the present embodiment includes four keys, respectively the first key, the second key, Third key and the 4th key, the first key are left button, and the second key is right key, and third key is middle key, and the 4th presses Key is OK key.Wherein, the first key includes pull-up resistor R4 and pull down resistor R11, pull-up resistor R4 connection 3.3V voltage, on One end of node connection key S2 between pull-up resistor R4 and pull down resistor R11, the other end ground connection of key S2, pull down resistor The other end of R11 connects main control chip U2, and when pressing the button S2, pull down resistor R11 only has 1K, and the first key exports low level, When unclamping key S2, output is pulled up resistance R4 and pull down resistor R11 pull-up, and the first key exports high level；Second key packet Include pull-up resistor R6 and pull down resistor R19, pull-up resistor R6 connection 3.3V voltage, between pull-up resistor R6 and pull down resistor R19 Node connection key S3 one end, the other end ground connection of key S3, the other end of pull down resistor R19 connects main control chip U2, When pressing the button S3, the second key exports low level, and when unclamping key S3, output is pulled up on resistance R6 and pull down resistor R19 It draws, the second key exports high level；Third key includes pull-up resistor R8 and pull down resistor R20, pull-up resistor R8 connection 3.3V Voltage, one end of the node connection key S4 between pull-up resistor R8 and pull down resistor R20, the other end ground connection of key S4, under The other end of pull-up resistor R20 connects main control chip U2, and when pressing the button S4, third key exports low level, when unclamping key S4, Output is pulled up resistance R8 and pull down resistor R20 pull-up, and third key exports high level；4th key includes pull-up resistor R10 With pull down resistor R26, pull-up resistor R10 connection 3.3V voltage, the node connection between pull-up resistor R10 and pull down resistor R26 One end of key S5, the other end ground connection of key S5, the other end of pull down resistor R26 connect main control chip U2, when pressing the button S5, the 4th key export low level, and when unclamping key S5, output is pulled up resistance R10 and pull down resistor R26 pull-up, and the 4th presses Key exports high level.

Main control chip U2 is also connected with memory, and memory uses SPI-FLASH device, for storing parameter, including it is humorous Vibration parameter, setting parameter etc..

Main control chip U2 is also connected with usb interface unit, and usb interface unit includes the USB core connecting with main control chip U2 The model USBLC6 of piece ESD1 and interface J7, the USB chip ESD1 being connect with USB chip ESD1, the model of interface J7 SIP254。

Main control chip U2 is also connected with touch unit, and touch unit includes the partial pressure electricity being connected between 3.3V voltage and ground R99 and R101 is hindered, is connected between the resistance R100 and base stage connection resistance R99 and R101 of the intermediate node of resistance R99 and R101 Divider resistance R98 and R96, partial pressure electricity are in series between the collector and 3.3V voltage of the triode Q10 of node, triode Q10 The grid of the node connection metal-oxide-semiconductor Q9 between R98 and R96 is hindered, the source electrode of metal-oxide-semiconductor Q9 connects 3.3V voltage, the drain electrode of metal-oxide-semiconductor Q9 Connection touches interface J1, touches interface J1 and connect with main control chip U2, touches interface J1 and is connected with touch tablet, main control chip U2 hair The power supply enable signal TP_PWEN sent passes through the base stage of resistance R100 to triode Q10, when power supply enable signal TP_PWEN is H When, triode Q10 conducting, metal-oxide-semiconductor Q9 conducting.

Main control chip U2 is also connected with external interface J4.

Main control chip U2 is also connected with buzzer.

Embodiment 3:

The present embodiment provides a kind of extracting solutions, the guanidine hydrochloride of guanidine thiocyanate and 1mol/L including 1.6mol/L, and pass through The pH to 4 of acetic acid adjusting extracting solution.The extracting solution is applied to and is used to save tissue or cell.

Embodiment 4:

The present embodiment provides a kind of extracting solutions, comprising:

Wherein, this extracting solution passes through second acid for adjusting pH to 4.The extracting solution is applied in extracting RNA.

Embodiment 5:

The present embodiment provides a kind of extracting solutions, including extracting solution A and extracting solution B；

Wherein extracting solution A includes the guanidine thiocyanate of 1.6mol/L and the guanidine hydrochloride of 1mol/L；

And extracting solution B includes the thiocyanic acid of the water saturation phenol of 57vt%, the guanidine thiocyanate of 0.24mol/L, 0.6mol/L The SDS of ammonium, the glycerol of 11.25vt% and 0.01mol/L；Surplus is solvent.

The pH of the extracting solution A and extracting solution B is 4.

The RNA amount extracted using the extracting solution in the present embodiment is measured A260/A280 and existed in 1000-1800 μ g/mL 1.6-1.9 range, A260/A230 is in 1.6-1.9 range.Agarose gel electrophoresis figure piece is as shown in Figure 7.

Using said extracted liquid A and extracting solution B and utilize the animals and plants of embodiment 1 or embodiment 2 based on ultrasonic principle The method of ablation of tissue instrument rapidly extracting RNA, comprising steps of 1. first save tissue or cell, the extracting solution A with extracting solution A The guanidine hydrochloride of guanidine thiocyanate and 0.5-1.5mol/L including 1.2-2.0mol/L, and pH is 4；

2. extracting solution B is added in the tissue or cell that save again to extracting solution A, it is mixed by inversion, obtains mixture, it is described to mention Taking liquid B includes with the water saturation phenol of the 30-65vt% of volume fraction and the glycerol of 9-15vt%, and surplus is solvent；

3. extract RNA from tissue, the mixture that step 2 obtains is disappeared using the animal vegetable tissue based on ultrasonic principle Melt instrument and carries out ultrasonication；Operation when extracting RNA from cell, directly in progress following step 5；

4. the mixture after ultrasonication is mixed by inversion, wherein ultrasonic power is 0.5-1000W, ultrasonic time 0.5- 30s；

5. chloroform is added, is centrifuged after mixing from the mixture of cell extraction RNA or the mixture of step 4 in step 3, Take supernatant liquid；

6. isopropanol is added in supernatant liquid, it is centrifuged after mixing, discards liquid, obtains sediment；

7. ethanol solution is added into sediment, it is centrifuged after mixing, discards liquid；

8. water dissolution is added after ethyl alcohol volatilization.

Experiment 1:

8 1.5mlEP pipes are taken, each EP pipe is added 4mg rat liver tissue, each EP pipe is numbered, wherein 1, 3,5, No. 7 EP pipes are separately added into 250ul extracting solution A, save 48h at room temperature, and 2,4,6, No. 8 EP pipes are separately added into 250ulRNAlater solution, saves 48h at room temperature.

750ul extracting solution B is separately added into above-mentioned all EP pipes again.

Supersonic melting, ultrasonic time 5s, frequency 20kHz are carried out after above-mentioned 1-8 EP pipe is mixed by inversion, power is 100W；EP pipe after ultrasound is reverse several times, mix mixture therein；To being separately added into 200ul in treated EP pipe Chloroform after being acutely mixed by inversion 30s, is centrifuged 1min under the centrifugal force of 10000g, takes supernatant liquid；By the supernatant liquid of acquisition It is poured into the clean EP pipe of another batch of reference numeral respectively, is then respectively adding 500ul isopropanol, after being slowly mixed by inversion 50s, It is centrifuged 1min under the centrifugal force of 10000g, discards liquid, obtains sediment；It is separately added into above-mentioned sediment 600ul75vt% ethanol solution after being slowly mixed by inversion 30s, is centrifuged 2min under the centrifugal force of 10000g, discards liquid；It will EP pipe be placed in after air drying 5min 40ulddH2O dissolution be added；The RNA of acquisition is subjected to micro ultraviolet specrophotometer Measure OD value and agarose gel electrophoresis detection.

It obtains 1 data of experiment and carries out interpretation of result, obtain the provable use extracting solution A of result compared to existing RNAlater solution can obtain the RNA DNA purity of better quality.

This uses A liquid of the invention, increases and extend the tissue preserration time, using A liquid of the invention, with present city On face RNA extract reagent Trizol, preservation effect is compared, respectively using both the above reagent to Rats Spleen tissue into Row saves, and deposits in room temperature and 37 DEG C two kinds, and the holding time is arranged at 7 days or more, after tissue morphology after preservation is observed, It was found that Trizol save spleen tissue all dissolve, we invent A liquid save tissue it is normal, after to two kinds save spleen Dirty tissue carries out RNA extraction, and the RNA amount that as a result the A liquid of the present embodiment extracts is enough to be higher by 3 times of water that Trizol saves tissue More than flat.

It extracts result after the spleen tissue that the present invention is saved with Trizol to compare, the tissue after as a result trizol is saved The RNA amount extracted measures A260/A280 about 1.583, A260/A230 is about 2.563 in 474 μ g/mL.Use this The RNA amount that the RNAlater of invention is extracted measures A260/A280 about 1.8, A260/A230 is 2.2 in 1241 μ g/mL Up and down.

Experiment 2:

8 1.5mlEP pipes are equally taken, each EP pipe is added 4mg rat liver tissue, each EP pipe is numbered, In, all EP pipes are separately added into 250ulRNAlater solution, save 48h at room temperature.

750ul extracting solution B is separately added into above-mentioned 1,3,5, No. 7 EP pipe therein again；And divide to 2,4,6, No. 8 EP pipes Not Jia Ru 1ml TRIzol solution.

It obtains 1 data of experiment and carries out interpretation of result, obtain the provable use extracting solution B of result compared to existing TRIzol solution can obtain the RNA DNA purity of better quality.

The RNA that the present embodiment extracting solution A is extracted after mixing with extracting solution B is extracted more compared to existing Trizol solution The RNA of good quality carries out RNA extraction to Rats Spleen tissue using both the above reagent, carries out concentration survey to the RNA after extraction Fixed, the RNA amount that Trizol is extracted is typically in 400 μ g/mL, while being measured to A260/A230, and measured value generally exists 1.5 horizontal.A260/A280 measurement result is in 1.8 levels.And the RNA amount that the reagent of the present embodiment extracts 1200 μ g/mL with Upper level, while A260/A230 is measured, measured value is generally in 1.8 levels.A260/A280 measurement result is in 1.7 water It is flat.

Experiment 3:

750ul extracting solution B is separately added into above-mentioned 1,3,5, No. 7 EP pipe；It discards in above-mentioned 2,4,6, No. 8 EP pipes RNA later solution is added after 1 × PBS is washed 3 times and abandons liquid, then is separately added into the TRIzol solution of 1ml.

Lysate B mixed mode is added using the preservation A liquid of the present embodiment, the RNA DNA purity extracted is high, using 1% The RNA that agarose gel electrophoresis extracts our reagent is confirmed, the RNA mass and effect that the present embodiment reagent extracts It is better than the RNA that trizol is extracted.The two comparison result is that the RNA amount that Trizol is extracted is typically in 400 μ g/mL, together When A260/A230 is measured, measured value generally 1.5 level.A260/A280 measurement result is in 1.8 levels.And this implementation The RNA amount that the reagent of example extracts is measured A260/A230 in 1200 μ g/mL level above, and measured value generally exists 1.8 horizontal.A260/A280 measurement result is in 1.7 levels.

Experiment 4:

6 1.5mlEP pipes are taken, each EP pipe is added about 106 Panc-1 cells, each EP pipe is numbered, wherein 1,2, No. 3 EP pipes are separately added into 250ul extracting solution A and 750ul extracting solution B, and 4,5, No. 6 EP pipes are separately added into 1mlTRIzol；It will Above-mentioned EP pipe is reverse several times, mixes mixture therein；It is separately added into 200ul chloroform in above-mentioned EP pipe, it is acutely reverse mixed After even 10s, it is centrifuged 1min under the centrifugal force of 10000g, takes supernatant liquid；

The supernatant liquid of acquisition is poured into respectively in the clean EP pipe of another batch of reference numeral, 500ul is then respectively adding Isopropanol after being slowly mixed by inversion 50s, is centrifuged 1min under the centrifugal force of 10000g, discards liquid, obtains sediment；Upwards State and be separately added into 600ul75vt% ethanol solution in sediment, after being slowly mixed by inversion 30s, under the centrifugal force of 10000g from Heart 1min, discards liquid；EP pipe is placed in after air drying 5min, 40ulddH2O dissolution is added；The RNA of acquisition is carried out micro- Measure ultraviolet specrophotometer measurement OD value and agarose gel electrophoresis detection.

Using the agent prescription of the present embodiment, RNA extraction and quality and Purity assessment are carried out, from spectrophotometric determination Result and agarose gel electrophoresis results be compared, the RNA amount that the present embodiment result is extracted is in the 1200 above water of μ g/mL It is flat, while A260/A230 is measured, measured value is generally in 1.8 levels.A260/A280 measurement result is in 1.7 levels.

Embodiment 6:

And extracting solution B includes the thiocyanic acid of the water saturation phenol of 57vt%, the guanidine thiocyanate of 0.24mol/L, 0.6mol/L Ammonium, the glycerol of 11.25vt%, the beta -mercaptoethanol of SDS, 0.1wt% of 0.3wt%, 0.08mmol/L potassium citrate, The bromophenol blue of 0.1wt ‰, the sodium acetate of 0.15mol/L, the isopropanol of 8vt%, 0.1wt% 8-hydroxyquinoline and 0.1mmol/ The EDTA of L.

The pH of the extracting solution A and extracting solution B is adjusted to 4 with acetic acid.

And it is directed to the above-mentioned best approach, is tested in a manner of variable by setting multiple groups contrast groups by controlling.

Experimental group 1:

The present embodiment provides a kind of extracting solutions, including extracting solution A and extracting solution B；Wherein extracting solution A includes 1.6mol/L's The guanidine hydrochloride of guanidine thiocyanate and 1mol/L；

And extracting solution B include the water saturation phenol of 57vt%, the ammonium thiocyanate of 0.6mol/L, 11.25vt% glycerol, The beta -mercaptoethanol of SDS, 0.1wt% of 0.3wt%, the potassium citrate of 0.08mmol/L, 0.1wt ‰ bromophenol blue, The sodium acetate of 0.15mol/L, the isopropanol of 8vt%, the 8-hydroxyquinoline of 0.1wt% and the EDTA of 0.1mmol/L.

Experimental group 2:

And extracting solution B includes the thiocyanic acid of the water saturation phenol of 57vt%, the guanidine thiocyanate of 0.24mol/L, 0.6mol/L Ammonium, the glycerol of 11.25vt%, the beta -mercaptoethanol of 0.1wt%, the potassium citrate of 0.08mmol/L, 0.1wt ‰ bromophenol blue, The sodium acetate of 0.15mol/L, the isopropanol of 8vt%, the 8-hydroxyquinoline of 0.1wt% and the EDTA of 0.1mmol/L.

Experimental group 3:

The pH of the extracting solution A and extracting solution B is 7 or 2.

Experimental group 4:

And extracting solution B includes the thiocyanic acid of the water saturation phenol of 57vt%, the guanidine thiocyanate of 0.24mol/L, 0.6mol/L Ammonium, the glycerol of 11.25vt%, the potassium citrate of SDS, 0.08mmol/L of 0.3wt%, the bromophenol blue of 0.1wt ‰, 0.15mol/ The sodium acetate of L, the isopropanol of 8vt%, the 8-hydroxyquinoline of 0.1wt% and the EDTA of 0.1mmol/L.

The pH of the extracting solution A and extracting solution B passes through acetic acid and is adjusted to 4.

Experimental group 5:

And extracting solution B includes the thiocyanic acid of the water saturation phenol of 57vt%, the guanidine thiocyanate of 0.24mol/L, 0.6mol/L Ammonium, the glycerol of 11.25vt%, the beta -mercaptoethanol of SDS, 0.1wt% of 0.3wt%, the bromophenol blue of 0.1wt ‰, 0.15mol/L Sodium acetate, the isopropanol of 8vt%, the 8-hydroxyquinoline of 0.1wt% and 0.1mmol/L EDTA.

Experimental group 6:

And extracting solution B includes the thiocyanic acid of the water saturation phenol of 57vt%, the guanidine thiocyanate of 0.24mol/L, 0.6mol/L Ammonium, the glycerol of 11.25vt%, the potassium citrate of SDS, 0.08mmol/L of 0.3wt%, the bromophenol blue of 0.1wt ‰, 0.15mol/ The EDTA of the sodium acetate of L, the isopropanol of 8vt% and 0.1mmol/L.

Experimental group 7:

And extracting solution B includes the thiocyanic acid of the water saturation phenol of 57vt%, the guanidine thiocyanate of 0.24mol/L, 0.6mol/L Ammonium, the glycerol of 11.25vt%, the beta -mercaptoethanol of SDS, 0.1wt% of 0.3wt%, 0.08mmol/L potassium citrate, The bromophenol blue of 0.1wt ‰, the isopropanol of 8vt%, the 8-hydroxyquinoline of 0.1wt% and the EDTA of 0.1mmol/L.

Experimental group 8:

And extracting solution B includes the thiocyanic acid of the water saturation phenol of 57vt%, the guanidine thiocyanate of 0.24mol/L, 0.6mol/L Ammonium, the glycerol of 11.25vt%, the beta -mercaptoethanol of SDS, 0.1wt% of 0.3wt%, 0.08mmol/L potassium citrate, The bromophenol blue of 0.1wt ‰, the sodium acetate of 0.15mol/L, the isopropanol of 8vt% and the 8-hydroxyquinoline of 0.1wt%.

The pH of extracting solution A and extracting solution B pass through acetic acid and are adjusted to 4.

Experimental group 9:

And extracting solution B include the water saturation phenol of 57vt%, 1mmol/L DTT, 0.24mol/L guanidine thiocyanate, The ammonium thiocyanate of 0.6mol/L, the glycerol of 11.25vt%, 0.3wt% SDS, 0.1wt% beta -mercaptoethanol, 0.08mmol/ The potassium citrate of L, the bromophenol blue of 0.1wt ‰, the urea of 5mol/L, the sodium acetate of 0.15mol/L, 8vt% isopropanol, The 8-hydroxyquinoline of 0.1wt% and the EDTA of 0.1mmol/L.

Unified experimental verification is carried out for the above-mentioned experimental group for lacking and increasing different component, every group takes 8 1.5mlEP pipe, each EP pipe are added 4mg rat liver tissue, each EP pipe are numbered, wherein all EP pipes add respectively Enter 250ul extracting solution A, saves 48h respectively at room temperature.

And 750ul extracting solution B is separately added into above-mentioned all EP pipes.

Interpretation of result is as follows: experimental group 1 because lack guanidine thiocyanate directly result in cracking it is insufficient, influence RNA extraction Constant.The RNA amount extracted significantly reduces, and extracts Comparative result with original formulation, lacks the guanidine thiocyanate reagent of 0.24mol/L The RNA amount extracted is on the low side, and the RNA amount extracted will be above or below 300 μ g/mL, and A260/A230 is about 1.7, A260/A280 It 2 or more, is quantitative determined using spectrophotometer, the RNA purity extracted is sentenced using agarose gel electrophoresis It is fixed.Cracking is insufficient because the SDS for lacking 0.3wt% is directly resulted in for experimental group 2, influences the constant of RNA extraction.It extracts RNA amount significantly reduces, and extracts Comparative result with original formulation, and it is on the low side to lack the RNA amount that the SDS reagent of 0.3wt% extracts, SDS Directly affect cracking, quantitative determined using spectrophotometer, using agarose gel electrophoresis to the RNA purity extracted into Row determines that the yield of RNA is above or below 200 μ g/mL.

Experimental group 3 had both influenced RNA yield because pH value changes, and it is impure to also result in the RNA extracted, introduced gene Group pollution.The RNA purity extracted is compared using agarose gel electrophoresis, it can be found that changing the reagent of pH value, There is genome pollution.PH value changes, and directly will lead to genome pollution, will not influence the yield of RNA.Experimental group 4 is because of β-mercapto Base ethyl alcohol is as reducing agent, it is ensured that welcomes the ammonium sulfate in liquid not oxidized, guarantees reagent stability, lack the β-of 0.1wt% Mercaptoethanol reagent will result directly in reagent save it is unstable, the resting period will lead to reagent color long afterwards and change.Experiment Group 5 is because lack the potassium citrate of 0.08mmol/L, it will reagent buffer system is influenced, to influence experimental result.And it is real It tests group 6 and lacks 8-hydroxyquinoline, because 8-hydroxyquinoline effect is to maintain the stability of water-saturated phenol, prevent the oxygen of water-saturated phenol Change.Influence B liquid preservation and stability for a long time.Because increasing sugar pollution in experimental group 7, influences RNA and extract quality.And it tests Lack EDTA described in group 8 and will increase and extracts the risk that RNA is degraded by RNase.Because the urea of 5mol/L and 1mmol/L's DTT is the effect for inhibiting RNase, so that the RNA extracted is from degradation.Therefore the RNA saved in experimental group 9 has dropped Solution.

The present invention is not limited to above-mentioned optional embodiment, anyone can show that other are various under the inspiration of the present invention The product of form, however, make any variation in its shape or structure, it is all to fall into the claims in the present invention confining spectrum Technical solution, be within the scope of the present invention.

Claims

1. utilizing the method for the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle, it is characterised in that: including step It is rapid:

G1. tissue first is saved with extracting solution A or cell, the extracting solution A include the guanidine thiocyanate and 0.5- of 1.2-2.0mol/L The guanidine hydrochloride of 1.5mol/L, and pH is 4；

G2. extracting solution B is added in the tissue or cell saved again to extracting solution A, is mixed by inversion, obtains mixture, the extraction Liquid B includes with the water saturation phenol of the 30-65vt% of volume fraction and the glycerol of 9-15vt%, and surplus is solvent；

Extracting solution B further include with the guanidine thiocyanate of the 0.20-1mol/L of molar concentration meter, 0.4-0.8mol/L ammonium thiocyanate and The surfactant of 0.0073-0.018mol/L, and pH is adjusted to 4；

When G3. extracting RNA from tissue, the mixture that step G2 is obtained is melted using the animal vegetable tissue based on ultrasonic principle Instrument carries out ultrasonication；Operation when extracting RNA from cell, in directly progress following step G5；

G4. the mixture after ultrasonication is mixed by inversion；

G5. chloroform is added, is centrifuged, takes after mixing from the mixture of the mixture of cell extraction RNA or step G4 in step G3 Supernatant liquid；

G8. water dissolution is added after ethyl alcohol volatilization；

The animal vegetable tissue ablatograph based on ultrasonic principle, comprising: power supply, DCDC converting unit, main control unit, DCDC function Rate adjusts unit, driving unit, voltage transformation unit, resonant element, ultrasonic transducer and sampling unit；

The power supply, by making after DCDC converting unit and DCDC power adjustment unit conversion voltage to animal vegetable tissue ablatograph With；Main control unit, output the first pwm signal control DCDC power adjustment unit exports adjustable voltage, and passes through acquisition transformation list The feedback of the sampling unit of the output end loop voltage and electric current of member, the second pwm signal for exporting two-way complementary duty cycle extremely drive Moving cell simultaneously passes through voltage transformation unit transformation, then adjusts resonance by the resonant element being set in the output end loop of voltage transformation unit Point arrives the resonance point of ultrasonic vibrator, so that the ultrasonic vibrator being set in the output end loop of voltage transformation unit works in resonant state.

2. the method according to claim 1 using the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle, It is characterized by: the surfactant in the extracting solution B is lauryl sodium sulfate.

3. the method according to claim 1 using the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle, It is characterized by: the extracting solution A uses salt acid for adjusting pH value, the extracting solution B uses second acid for adjusting pH value.

4. animal vegetable tissue ablatograph rapidly extracting of the utilization based on ultrasonic principle according to claim 1 to 3 The method of RNA, it is characterised in that: further include β-mercapto that molar concentration is 6.4mmol/L-19.20mmol/L in the extracting solution B Base ethyl alcohol.

5. animal vegetable tissue ablatograph rapidly extracting of the utilization based on ultrasonic principle according to claim 1 to 3 The method of RNA, it is characterised in that: further include the potassium citrate that molar concentration is 0.05-0.15mmol/L in the extracting solution B.

6. animal vegetable tissue ablatograph rapidly extracting of the utilization based on ultrasonic principle according to claim 1 to 3 The method of RNA, it is characterised in that: further include in the extracting solution B molar concentration be 74.62 μm of ol/L-223.87 μm of ol/L Bromophenol blue.

7. animal vegetable tissue ablatograph rapidly extracting of the utilization based on ultrasonic principle according to claim 1 to 3 The method of RNA, which is characterized in that further include the sodium acetate that molar concentration is 0.01-2mol/L in the extracting solution B.

8. animal vegetable tissue ablatograph rapidly extracting of the utilization based on ultrasonic principle according to claim 1 to 3 The method of RNA, it is characterised in that: further include the isopropanol that volume fraction is 5-25% in the extracting solution B.

9. animal vegetable tissue ablatograph rapidly extracting of the utilization based on ultrasonic principle according to claim 1 to 3 The method of RNA, it is characterised in that: further include the 8- that molar concentration is 0.689mmol/L-68.9mmol/L in the extracting solution B Oxyquinoline.

10. animal vegetable tissue ablatograph of the utilization based on ultrasonic principle according to claim 1 to 3 quickly mentions The method for taking RNA, it is characterised in that: further include the DTT that molar concentration is 0.5-1000mmol/L in the extracting solution B.

11. animal vegetable tissue ablatograph of the utilization based on ultrasonic principle according to claim 1 to 3 quickly mentions The method for taking RNA, it is characterised in that: further include the urea that molar concentration is 0.5-5mol/L in the extracting solution B.

12. animal vegetable tissue ablatograph of the utilization based on ultrasonic principle according to claim 1 to 3 quickly mentions The method for taking RNA, it is characterised in that: further include the EDTA that molar concentration is 0.1-500mmol/L in the extracting solution B.

13. animal vegetable tissue ablatograph of the utilization based on ultrasonic principle according to claim 1 to 3 quickly mentions The method for taking RNA, it is characterised in that: further include the ammonium acetate that molar concentration is 0.01-10mmol/L in the extracting solution B.

14. animal vegetable tissue ablatograph of the utilization based on ultrasonic principle according to claim 1 to 3 quickly mentions The method for taking RNA, it is characterised in that: the water in the step G8 is distilled water, deionized water or ultrapure water.

15. animal vegetable tissue ablatograph of the utilization based on ultrasonic principle according to claim 1 to 3 quickly mentions The method for taking RNA, it is characterised in that: the gross mass of the extracting solution A and extracting solution B is the 100-150 of tissue or cell quality Times.

16. animal vegetable tissue ablatograph of the utilization based on ultrasonic principle according to claim 1 to 3 quickly mentions The method for taking RNA, it is characterised in that: in the step G3, ultrasonic power 0.5-1000W, ultrasonic time 0.5-30s.

17. animal vegetable tissue ablatograph of the utilization based on ultrasonic principle according to claim 1 to 3 quickly mentions The method for taking RNA, it is characterised in that: in the step G5, the volume of chloroform is 0.1-0.4 times of volume of mixture.

18. animal vegetable tissue ablatograph of the utilization based on ultrasonic principle according to claim 1 to 3 quickly mentions The method for taking RNA, it is characterised in that: in the step G6, the volume ratio of isopropanol and supernatant liquid is 1:1.

19. the side according to claim 1 using the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle Method, it is characterised in that: the DCDC power adjustment unit of the animal vegetable tissue ablatograph includes: that switch control signal receiving end connects Connect the switch control signal reception circuit that main control unit receives switch control signal；

Power supply is converted to the 4th of adjustable voltage output by the switch control signal and the first pwm signal of main control unit transmission DCDC converting unit.

20. the side according to claim 19 using the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle Method, it is characterised in that: the 4th DCDC converting unit of the animal vegetable tissue ablatograph includes: successively to handle the of supply voltage The filtering of four voltage inputs, the 4th voltage conversion chip U1 and the filtering of the 4th voltage output, the filtering of the 4th voltage input include connection Pass through energy storage inductor L1 in the upper tube driving signal reference point pin SW of the filter capacitor of power end, the 4th voltage conversion chip U1 It is connected to adjustable voltage output end, the divider resistance R32 and R47 for being connected on adjustable voltage output end carry out voltage sample and be input to The upper tube driving signal reference point pin of the reference voltage pin FB of 4th voltage conversion chip U1, the 4th voltage conversion chip U1 SW is also connected with sustained diode 2, and enable pin EN is also connected with resistance R33, resistor timing/external clock pin RT/ CLK is also connected with frequency divider resistance R85, and frequency compensation pin COMP is also connected with for adjusting loop, burning voltage output Capacitor C51, C49 and resistance R46, wherein capacitor C49 is in parallel with concatenated capacitor C51 and resistance R46, the 4th voltage output Filtering includes the filter capacitor for being connected to adjustable voltage output end；

It includes: the resistance being series between converted voltage and switch control signal receiving end that switch control signal, which receives circuit, R104 and R105, emitter connects the node between converted voltage, base stage connection resistance R104 and R105 and collector connects The triode Q12 of the enable pin EN of 4th voltage conversion chip U1, when switch control signal is L, triode Q12 conducting, 4th voltage conversion chip U1 opens work；

It includes: RC filtering and the first voltage follower U3B for successively handling the first pwm signal that first pwm signal, which receives circuit, the Received first pwm signal in one pwm signal receiving end is input to first voltage follower U3B's after being filtered by RC filtering The output end of non-inverting input terminal, first voltage follower U3B is connected between divider resistance R32 and R47 by divider resistance R54 Node.

21. the side according to claim 1 using the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle Method, it is characterised in that: the driving unit of the animal vegetable tissue ablatograph includes the first driving unit and the second driving unit, the Second pwm signal receiving end N of one driving unit receives the second pwm signal of the first via that main control unit is sent, and the first driving is single The Same Name of Ends of the primary coil of the output end connection voltage transformation unit of member, the second pwm signal receiving end P of the second driving unit are received Second the second pwm signal of tunnel that main control unit is sent, the primary coil of the output end connection voltage transformation unit of the second driving unit Different name end, by the second pwm signal receiving end N received the second pwm signal of first via control the first driving metal-oxide-semiconductor Q6 conducting/ Cut-off is ended/is led by received the second driving of the second pwm signal of the second tunnel control metal-oxide-semiconductor Q2 of the second pwm signal receiving end P It is logical.

22. the side according to claim 21 using the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle Method, it is characterised in that: the voltage transformation unit of the animal vegetable tissue ablatograph be push-pull transformer, DCDC power adjustment unit can Adjust the different name end of the primary side first coil of voltage output end connection push-pull transformer and the Same Name of Ends of the second coil；

First driving unit includes the resistance R10 and R14 being series between the second pwm signal receiving end N, and grid connects Node, source electrode ground connection and drain electrode between connecting resistance R10 and R14 are the output end of the first driving unit and connect push-pull transformer Primary side first coil Same Name of Ends first driving metal-oxide-semiconductor Q6；

Second driving unit includes the resistance R5 and R13 being series between the second pwm signal receiving end P, and grid connects Node, source electrode ground connection and drain electrode between connecting resistance R5 and R13 are the output end of the second driving unit and connect push-pull transformer The second coil of primary side different name end second driving metal-oxide-semiconductor Q2.

23. the side according to claim 1 using the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle Method, it is characterised in that: the resonant element of the animal vegetable tissue ablatograph is LC series resonance, and LC series resonance includes being series at Inductance T1 and capacitor C1 in the output end loop of voltage transformation unit.

24. the side according to claim 1 using the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle Method, it is characterised in that: the sampling unit of the animal vegetable tissue ablatograph includes the output end loop voltage for acquiring voltage transformation unit Voltage sampling unit and acquisition voltage transformation unit output end loop current current sampling unit, correspondingly, voltage transformation unit It exports end loop and is equipped with multiple concatenated sampling resistors；Voltage sampling port by being connected to the intermediate node of multiple sampling resistors is adopted Collecting voltage signal is then forwarded to voltage sampling unit and is filtered and amplifies, and is then forwarded to main control unit and does resonance adjusting； Current sampling port by being connected on the output end loop of voltage transformation unit acquires current signal and is sent to current sampling unit progress Filtering and amplification, are then forwarded to main control unit and do resonance adjusting.

25. the side according to claim 1 using the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle Method, it is characterised in that: the main control unit of the animal vegetable tissue ablatograph includes main control chip U2 and main control chip peripheral circuit, Main control chip U2 exports the first pwm signal control DCDC power adjustment unit and exports adjustable voltage, and passes through acquisition voltage transformation unit Output end loop voltage and electric current sampling unit feedback, export two-way complementary duty cycle the second pwm signal to drive Unit.

26. the side according to claim 25 using the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle Method, it is characterised in that: the main control unit further includes companion chip U1 and companion chip peripheral circuit, the master control core of main control unit Piece U2 is referred to by the feedback of the output end loop voltage of acquisition voltage transformation unit and the sampling unit of electric current to companion chip U1 transmission It enabling, companion chip U1 receives the instruction of main control chip U2, export the second pwm signal of two-way complementary duty cycle to driving unit, Meanwhile the second pwm signal of two-way complementary duty cycle also feeds back to main control chip U2.

27. the side according to claim 25 using the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle Method, it is characterised in that: the second pwm signal of the two-way complementary duty cycle also respectively by the first signal amplification output circuit and Second signal amplification output circuit is sent to driving unit after amplifying.

28. the side according to claim 25 using the animal vegetable tissue ablatograph rapidly extracting RNA based on ultrasonic principle Method, it is characterised in that: the animal vegetable tissue ablatograph further includes the display unit, key, storage connecting with main control chip U2 Device, usb interface unit or/and touch unit.