CN108004324A - One kind is used for tumour cell nucleic acid aptamers screening circular card box device - Google Patents
One kind is used for tumour cell nucleic acid aptamers screening circular card box device Download PDFInfo
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- CN108004324A CN108004324A CN201711472080.0A CN201711472080A CN108004324A CN 108004324 A CN108004324 A CN 108004324A CN 201711472080 A CN201711472080 A CN 201711472080A CN 108004324 A CN108004324 A CN 108004324A
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- 238000012216 screening Methods 0.000 title claims abstract description 34
- 108091008104 nucleic acid aptamers Proteins 0.000 title claims abstract description 21
- 210000004881 tumor cell Anatomy 0.000 title claims abstract description 20
- 108091023037 Aptamer Proteins 0.000 claims abstract description 39
- 238000003860 storage Methods 0.000 claims abstract description 38
- 238000003825 pressing Methods 0.000 claims abstract description 36
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 29
- 239000007788 liquid Substances 0.000 claims description 42
- 239000000243 solution Substances 0.000 claims description 26
- 238000012546 transfer Methods 0.000 claims description 17
- 238000004140 cleaning Methods 0.000 claims description 15
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 12
- 108010019160 Pancreatin Proteins 0.000 claims description 11
- 239000007984 Tris EDTA buffer Substances 0.000 claims description 11
- 229940055695 pancreatin Drugs 0.000 claims description 11
- 230000001079 digestive effect Effects 0.000 claims description 10
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 10
- 239000002699 waste material Substances 0.000 claims description 9
- 229910052742 iron Inorganic materials 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 4
- 239000007799 cork Substances 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims 2
- 238000000034 method Methods 0.000 abstract description 8
- 238000012864 cross contamination Methods 0.000 abstract description 7
- 229920001971 elastomer Polymers 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- 108020004414 DNA Proteins 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 230000000994 depressogenic effect Effects 0.000 description 4
- 230000003321 amplification Effects 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 239000003446 ligand Substances 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 150000007523 nucleic acids Chemical class 0.000 description 3
- 238000012408 PCR amplification Methods 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 108020004682 Single-Stranded DNA Proteins 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000007846 asymmetric PCR Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 208000002925 dental caries Diseases 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000010079 rubber tapping Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 229920002379 silicone rubber Polymers 0.000 description 1
- 238000012549 training Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
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Abstract
The invention discloses one kind to be used for tumour cell nucleic acid aptamers screening cartridge device, including cartridge bodies and the cartridge reagent storage disk positioned at cartridge bodies bottom；Turntable, pressing plate and liquid-transfering gun point are provided with cartridge bodies, is provided with air-path interface and air filter film on the shell of cartridge bodies, air-path interface is connected with the side air flue interface of liquid-transfering gun point；Extended at the top of turntable and the first control interface is formed outside cartridge bodies, liquid-transfering gun point is fixed on turntable；Pressing plate is located above turntable, is extended at the top of pressing plate and the second control interface is formed outside cartridge bodies；Tissue Culture Dish adapter and reagent storage reacting hole position are provided with cartridge reagent storage disk.The closed cartridge device of the present invention, the problem of assembling is simple, available for automation mechanized operation, and whole aptamer screening process all carries out in closed cartridge, avoids cross contamination, a wheel or multi-turns screen process for aptamer can be completed, greatly improves the efficiency of aptamer screening.
Description
Technical field
The invention belongs to biology field, and in particular to one kind is used for tumour cell nucleic acid aptamers screening circular card
Box device, suitable for the research experiment screened to tumour cell nucleic acid aptamers, simultaneously can be used for other species nucleic acid
Aptamers screening test, such as albumen, excretion body.
Background technology
Aptamer is a bit of oligonucleotide sequence obtained through in-vitro screening, and high parent can be carried out with corresponding ligand
With power and strong specific combination, its appearance for chemical-biological educational circles and biomedical boundary provide it is a kind of it is new it is efficient quickly
The research platform of identification, and good application prospect is illustrated in many aspects.
At present, the screening of tumour cell nucleic acid aptamers is carried out mainly by having been manually done, and inefficiency, is fitted from screening nucleic acid
Ligand screening starts to being fully completed, it is necessary to which carrying out 20 takes turns left and right experiment, and whole process needs half a year to 1 year, wherein most heavy
Will the problem of be that cross contamination occurs, be mainly derived from PCR, aptamers transfer etc., once pollute, this wheel or even whole
Error just occurs in a the selection result, it is necessary to re-starts screening.At present, the method for cross contamination is avoided mainly by will be every
A step is divided among different experiments room and is operated, and laboratory is aerated, is sterilized, and this mode needs to take larger ground
Side arrangement laboratory, and it is more demanding to laboratory technician, even if in this way, the aerial nucleic acid times that is scattered can not so eliminate, in fact
In the experiment on border, cross-contamination issue exists always.
The content of the invention
Goal of the invention：In view of the above-mentioned problems, the present invention proposes that a kind of tumour cell nucleic acid aptamers that are used for screen circular card
Box device, the cartridge device are closed, assemble simply, can operate liquid relief, PCR reactions, aptamers transfer etc. at one
Carried out in the cartridge of closing, avoid cross-contamination issue, and can realize automation mechanized operation.
Technical solution：To achieve these goals, a kind of tumour cell nucleic acid aptamers that are used for are screened as representative of the present invention
Cartridge device, including cartridge bodies and the cartridge reagent storage disk positioned at cartridge bodies bottom；It is provided with the cartridge bodies
Turntable, pressing plate and liquid-transfering gun are sharp, and air-path interface and air filter film, air-path interface and liquid-transfering gun are provided with the shell of cartridge bodies
The side air flue interface connection of point；Extended at the top of the turntable and the first control interface, liquid-transfering gun are formed outside cartridge bodies
Point is fixed on turntable；The pressing plate is located above turntable, and the control of formation second outside cartridge bodies is extended at the top of pressing plate and is connect
Mouthful；Tissue Culture Dish adapter and reagent storage reacting hole position are provided with the cartridge reagent storage disk.
Preferably, the turntable bottom is circular thin panel, the raceway groove protruded by cartridge bodies interior is consolidated
It is fixed, there is perforate on turntable, liquid-transfering gun point is fixed by rifle point spring.
Preferably, the pressing plate bottom is cuboid thin plate, bottom is provided with rag iron spring.
Preferably, it is provided with air filtering core in the side air flue interface of the air-path interface and liquid-transfering gun point.It is fixed with
Air filtering core, filter core aperture are less than aptamers, prevent aptamers from spilling into pressure generator, produce pollution.
Further, the filter sizes of the air filter film are less than aptamers.The shell lateral plane part of cartridge bodies
Air filter film is fixed with, filter sizes are less than aptamers, are balanced for air pressure in keeping box and external pressure.
Wherein, the Tissue Culture Dish adapter is semiclosed bulge, and bottom is provided with shaft, can be with cartridge bodies
Part is tightly engaged into.
Preferably, the shaft is seamlessly connected with bulge, the two is to be connected as a single entity, and shaft one end projects to cartridge
The housing exterior of main body, forms shaft and protrudes control point.The head that shaft protrudes control point is linear type or other shapes interface,
Stir by hand or motor connecting interface rotates, control pivot, so as to control adapter to swing, cultivated for controlling
Liquid aggregation position in ware.
Further, the reagent storage reacting hole position includes the 9 holes position being successively set on cartridge reagent storage disk,
9 holes position is respectively library high temperature receiving hole, and library low-temp storage hole, with reference to liquid receiving hole, TE buffer solution receiving hole, gives up
Liquid receiving hole, cleaning solution receiving hole, pancreatin digestive juice receiving hole, PCR reacting holes, aptamers transfer receiving hole.
Preferably, the reagent storage reacting hole position is circular or square, bottom is closed with hemispherical or taper ending,
The orifice center point of hole position is distributed on the circumference that the center of circle is overlapped with the round turntable center of circle.
The library high temperature receiving hole and aptamers transfer receiving hole be detachable movable span position, the opening cork in hole
Closing, such as rubber stopper, silica gel plug.It can be separated from cartridge, for testing the transfer of aptamers before and after.
Operation principle：Carry out the first round aptamer screening start before, by DNA library be put into library high temperature receiving hole,
It is put into reference to liquid and is put into PCR reacting holes with reference to liquid receiving hole, PCR reaction reagents, TE buffer solutions is put into TE buffer solutions receiving hole, pancreas
Enzymic digestion liquid is put into pancreatin digestive juice receiving hole, cleaning solution is put into cleaning solution receiving hole, and Tissue Culture Dish is put into cell culture
Ware adapter, after completion, cartridge bodies and cartridge reagent storage disk are completed.Library high temperature receiving hole is put into 95 DEG C
After metal bath 5min, the first control interface at the top of turntable is rotated, the liquid-transfering gun point being fixed on turntable bottom disc is gone to
Directly over the high temperature receiving hole of library, the second control interface at the top of pressing plate is rotated, the cuboid thin plate of pressing plate bottom is gone into shifting
Directly over liquid rifle point, the second control interface at the top of pressing plate is firmly oppressed, pressing plate moves down, and liquid-transfering gun point is depressed into library high temperature storage
Inside hole, library is sucked by liquid-transfering gun point cavity by the plunger pump being connected with air-path interface, withdraws from and is applied at the top of pressing plate
Power, is acted on by rag iron spring elastic force, and pressing plate moves up, and since the elastic force of rifle point spring acts on, library height is moved on liquid-transfering gun point
Above gentle discharge hole, the first control interface at the top of turntable is rotated, the liquid-transfering gun point being fixed on turntable bottom disc is gone to
Directly over the low-temp storage hole of library, the second control interface at the top of pressing plate is rotated, the cuboid thin plate of pressing plate bottom is gone into shifting
Directly over liquid rifle point, interface at the top of pressing plate is firmly oppressed, pressing plate moves down, and liquid-transfering gun point is depressed into inside the low-temp storage hole of library,
Liquid-transfering gun point cavity Chinese library is put into the low-temp storage hole of library by plunger pump, after 0 DEG C of storage 10min, with liquid-transfering gun point
Library low-temp storage hole Chinese library is drawn to liquid receiving hole is combined, is mixed by liquid-transfering gun point piping and druming and combines liquid in liquid receiving hole
Afterwards, cleaning solution in cleaning solution receiving hole is drawn with liquid-transfering gun point to put into Tissue Culture Dish adapter in Tissue Culture Dish, gently
The shaft of left-right rotation Tissue Culture Dish adapter 28 uses liquid-transfering gun point to draw Tissue Culture Dish supernatant after protruding control point 2min,
Put to waste liquid receiving hole, after repeated washing 3 times, drawn to combine liquid in liquid receiving hole and move to Tissue Culture Dish with liquid-transfering gun point and fitted
In orchestration 28 in Tissue Culture Dish, dashed forward in 4 DEG C of environment by the shaft of motor gently left-right rotation Tissue Culture Dish adapter
Go out control point 2h, draw Tissue Culture Dish aqueous supernatant with liquid-transfering gun point, put to waste liquid receiving hole, cleaning is drawn with liquid-transfering gun point
Cleaning solution is put into Tissue Culture Dish adapter in Tissue Culture Dish in liquid receiving hole, and gently left-right rotation Tissue Culture Dish is adapted to
Tissue Culture Dish supernatant is drawn with liquid-transfering gun point after device 2min, is put to waste liquid receiving hole, after repeated washing 3 times, draws TE bufferings
TE buffer solutions are into Tissue Culture Dish in liquid receiving hole 32, then draw pancreatin in pancreatin digestive juice receiving hole with liquid-transfering gun point and digest
Liquid in Tissue Culture Dish is moved in PCR reacting holes, carried out after mixing to after Tissue Culture Dish 10min by liquid with liquid-transfering gun point
PCR amplification, after amplification, liquid-transfering gun point is drawn amplified production and is moved to above aptamers transfer receiving hole, is filled in through rubber
Entering in hole, and amplified production is discharged into hole, turn aptamers transfer receiving hole, it is removed from cartridge, so far, the first round
Aptamer screening is completed, and will disassemble aptamers transfer receiving hole, is directly installed on the library high temperature of another cartridge
Library high temperature receiving hole of the hole site as new cartridge is stored, directly can carry out the second wheel or the n-th wheel experiment by above-mentioned steps,
Until aptamer screening is completed.
Beneficial effect：Compared by the prior art, the invention has the advantages that：The present invention's fits for tumour cell nucleic acid
Ligand screening circular card box device be it is a kind of it is closed, assembling is simple, the cartridge device available for automation mechanized operation.Whole nucleic acid
Aptamers screening process all carries out in closed cartridge, wherein the PCR reactions for easily producing cross contamination occur in cartridge,
Aptamers library transfer between two cartridges is carried out by the reagent wells of closing, ensures that amplified production will not spill into air
In, the problem of avoiding cross contamination.Aptamer screening is carried out by cartridge at the same time to realize by self-reacting device, can
A wheel or multi-turns screen process for aptamer is completed, greatly improves the efficiency of aptamer screening.
Brief description of the drawings
Fig. 1 is cartridge device overall structure diagram of the present invention；
Fig. 2 is the structure diagram of cartridge bodies；
Fig. 3 is cartridge bodies side sectional view；
Fig. 4 is the reagent storage dish structure schematic diagram of cartridge device；
Fig. 5 is cartridge device entirety side sectional view.
Embodiment
Below in conjunction with drawings and examples, the invention will be further described.
Embodiment
As shown in Figs. 1-5, it is a kind of to be used for tumour cell nucleic acid aptamers screening cartridge device, including cartridge bodies 1 and position
Dismountable cartridge reagent storage disk 2 in 1 bottom of cartridge bodies；Turntable 21, pressing plate 22 and liquid relief are provided with cartridge bodies 1
Rifle point 25；The top of turntable 21 extends to the outside of cartridge bodies 1 and forms the first control interface 38, is protruding to the of the outside of shell
The head of one control interface is linear type or other shapes interface, stirs by hand or motor connects 38 turns of the first control interface
It is dynamic, dial movement is controlled, 21 bottom of turntable is circular thin panel, and the raceway groove protruded by 1 interior of cartridge bodies is fixed, and is turned
There are two perforates on disk 21, identical two liquid-transfering gun points 25 are fixed on turntable by perforate on turntable 21 and rifle point spring 26,
Prevent the pollution of reagent in pipetting processes, the rotational movement liquid-transfering gun point 25 of turntable 21 moves；Pressing plate 22 is located at turntable 21
Top, the top of pressing plate 22 extend to the outside of cartridge bodies 1 and form the second control interface 39；22 upper end of pressing plate is socketed on turntable 22
End, bottom is a cuboid or other shapes panel, and panel bottom is provided with rag iron spring 27, by the second of enclosure
39 applying power of control interface is oppressed or the second control interface 39 of rotary pressure plate causes lower end board to produce horizontal or vertical displacement,
It is bonded when pressing plate bottom is moved to 25 top of liquid-transfering gun point with 25 top of liquid-transfering gun point, pushes the second control interface 39 and transport downwards
When dynamic, compressing liquid-transfering gun point 25 moves downward, and after pressure revocation, pressing plate 22 passes through rag iron spring 27 respectively with liquid-transfering gun point 25
Set back with the spring force of rifle point spring 26.
Cartridge bodies 1 are cylinder, and the shell side of cartridge bodies 1 is made of arc surface and plane, outside cartridge bodies 1
There are one or more prominent air-path interfaces 23 in the plane of shell, air-path interface 23 connects plunger in 1 exterior portion of cartridge bodies
The air pump pressure generators such as pump, liquid-transfering gun is connected in 1 interior section of cartridge bodies by rubber tube or other materials pipeline
The side air flue interface of the side air flue interface of point 25, the suction tapping operation of control rifle point, air-path interface 23 and liquid-transfering gun point 25
Inside it is provided with air filtering core.Air filter film 24, the filter membrane hole of air filter film 24 are additionally provided with the plane of the shell of cartridge bodies 1
Footpath is less than aptamers.
Tissue Culture Dish adapter and reagent storage reacting hole position, Tissue Culture Dish are provided with cartridge reagent storage disk 2
Adapter is semiclosed bulge, and bottom is provided with shaft, and shaft is seamlessly connected with semiclosed bulge, one distal process of shaft
Go out to the housing exterior of cartridge bodies 1, form shaft and protrude control point 40, control is protruded by projecting to the shaft outside cartridge
The control Tissue Culture Dish adapter side-to-side movement of point 40, so as to drive Tissue Culture Dish side-to-side movement, reaches in Tissue Culture Dish
The purpose of portion's liquid accumulation.Reagent storage reacting hole position include be successively set on cartridge reagent storage disk 29 holes position, 8
Hold the hole position and 1 waste liquid hole position of reagent, 9 holes position respectively library high temperature receiving hole 29, library low-temp storage hole 30, knot
Conjunction liquid receiving hole 31, TE buffer solutions receiving hole 32, waste liquid receiving hole 33, cleaning solution receiving hole 34, pancreatin digestive juice receiving hole 35,
PCR reacting holes 36, aptamers transfer receiving hole 37.Each reagent storage reacting hole position to be circular or square, bottom with hemispherical or
Taper ending closure, the orifice center point of hole position are distributed on the circumference that the center of circle is overlapped with 22 center of circle of turntable.Wherein, library high temperature
Receiving hole 29 is detachable movable span position with aptamers transfer receiving hole 37, and the opening of hole position is closed with cork, such as rubber stopper, silicon
Rubber plug etc..25 bottoms of liquid-transfering gun point can wear out rubber stopper and inject liquid into the hole, which is isolated
Come, can store or be directly installed in another cartridge, realize the transfer of aptamers between two cartridges.
The present embodiment combination HepG2 cells carry out one wheel aptamer screening, key step have library processing, DNA with
Cell combination, cell separation and broken, aptamer amplification, aptamer shift.Used reagent has single stranded DNA text
Storehouse, with reference to liquid, cleaning solution, pancreatin digestive juice, TE buffer solutions, asymmetric PCR reaction reagent etc..
Before the aptamer screening of the progress first round starts, DNA library is put into library high temperature receiving hole 29, is put with reference to liquid
Enter to combine liquid receiving hole 31, PCR reaction reagents are put into PCR reacting holes 36, TE buffer solutions are put into TE buffer solutions receiving hole 32, pancreatin
Digestive juice is put into pancreatin digestive juice receiving hole 35, cleaning solution is put into cleaning solution receiving hole 34, and Tissue Culture Dish is put into cell training
Ware adapter 28 is supported, after completion, cartridge bodies 1 and cartridge reagent storage disk 2 are completed.By library high temperature receiving hole 29
After being put into 95 DEG C of metal bath 5min, first control interface 38 at the top of turntable 21 is rotated, will be fixed on 21 bottom disc of turntable
Liquid-transfering gun point 25 go to directly over library high temperature receiving hole 29, second control interface 39 at the top of pressing plate 22 is rotated, by pressing plate
The cuboid thin plate of 22 bottoms is gone to directly over liquid-transfering gun point 25, firmly oppresses the second control interface 39 at the top of pressing plate, pressing plate 22
Move down, liquid-transfering gun point 25 is depressed into inside the high temperature receiving hole of library, is inhaled library by the plunger pump being connected with air-path interface 23
Enter sharp 25 cavitys of liquid-transfering gun, withdraw from the power for being applied to second control interface 39 of the top of pressing plate 22, made by 27 elastic force of rag iron spring
With, pressing plate 22 moves up, and since the elastic force of rifle point spring 26 acts on, the top of library high temperature receiving hole 29 is moved on liquid-transfering gun point 25,
First control interface 38 at the top of turntable 21 is rotated, it is low that the liquid-transfering gun point 25 being fixed on 21 bottom disc of turntable is gone into library
Directly over gentle discharge hole 30, second control interface 39 at the top of pressing plate 22 is rotated, the cuboid thin plate of 22 bottom of pressing plate is gone to
Directly over liquid-transfering gun point 25, second control interface 39 of the top of pressing plate 22 is firmly oppressed, pressing plate 22 moves down, liquid-transfering gun point 25 is depressed into
The sharp 25 cavity Chinese libraries of liquid-transfering gun, are put into library low-temp storage hole 30,0 by the inside of library low-temp storage hole 30 by plunger pump
After DEG C storage 10min, 30 Chinese library of library low-temp storage hole is drawn to combining liquid receiving hole 31 with liquid-transfering gun point 25, passes through liquid relief
25 piping and druming of rifle point, which mixes, to be combined in liquid receiving hole 31 after liquid, and drawing cleaning solution in cleaning solution receiving hole 34 with liquid-transfering gun point 25 puts
Into Tissue Culture Dish adapter 28 in Tissue Culture Dish, gently the shaft of left-right rotation Tissue Culture Dish adapter 28 protrudes control
Tissue Culture Dish supernatant is drawn with liquid-transfering gun point 25 after 40,2min of system point, is put to waste liquid receiving hole 33, after repeated washing 3 times, is used
Liquid-transfering gun point 25 is drawn to be moved in Tissue Culture Dish adapter 28 in Tissue Culture Dish with reference to liquid in liquid receiving hole 31, at 4 DEG C
In environment by motor gently left-right rotation Tissue Culture Dish adapter 28 shaft protrude 40 2h of control point, with liquid-transfering gun point
25 draw Tissue Culture Dish aqueous supernatant, put to waste liquid receiving hole 33, are drawn with liquid-transfering gun point 25 clear in cleaning solution receiving hole 34
Washing lotion is put into Tissue Culture Dish adapter 28 in Tissue Culture Dish, is gently used after left-right rotation Tissue Culture Dish adapter 2min
Liquid-transfering gun point 25 draws Tissue Culture Dish supernatant, puts to waste liquid receiving hole 33, after repeated washing 3 times, draws the storage of TE buffer solutions
TE buffer solutions draw pancreatin digestive juice in pancreatin digestive juice receiving hole 35 into Tissue Culture Dish, then with liquid-transfering gun point 25 in hole 32
To Tissue Culture Dish 10min, liquid in Tissue Culture Dish is moved in PCR reacting holes 36 with liquid-transfering gun point 25, is mixed laggard
Row PCR amplification, after amplification, liquid-transfering gun point 25 draws amplified production and moves to 37 top of aptamers transfer receiving hole, through rubber
In rubber plug access aperture, and amplified production is discharged into hole, turn aptamers transfer receiving hole 37, it is removed from cartridge, extremely
This, the screening of first round aptamer is completed, and the aptamers disassembled the transfer receiving hole 37 in cartridge is directly installed on
Library high temperature receiving hole of the position of the library high temperature receiving hole 29 of another cartridge as new cartridge, can directly press above-mentioned step
It is rapid to carry out the second wheel or the n-th wheel experiment, until aptamer screening is completed.
Claims (10)
1. one kind is used for tumour cell nucleic acid aptamers screening cartridge device, it is characterised in that including cartridge bodies and positioned at card
The cartridge reagent storage disk of box main body bottom；Turntable, pressing plate and liquid-transfering gun point are provided with the cartridge bodies, cartridge bodies
Air-path interface and air filter film are provided with shell, air-path interface is connected with the side air flue interface of liquid-transfering gun point；The turntable
Top extend to the first control interface formed outside cartridge bodies, liquid-transfering gun point is fixed on turntable；The pressing plate, which is located at, to be turned
Above disk, extended at the top of pressing plate and the second control interface is formed outside cartridge bodies；It is provided with the cartridge reagent storage disk
Tissue Culture Dish adapter and reagent storage reacting hole position.
2. according to claim 1 be used for tumour cell nucleic acid aptamers screening cartridge device, it is characterised in that described
Turntable bottom is circular thin panel, and the raceway groove protruded by cartridge bodies interior is fixed, and has perforate on turntable, passes through rifle point
Spring fixes liquid-transfering gun point.
3. according to claim 1 be used for tumour cell nucleic acid aptamers screening cartridge device, it is characterised in that the pressure
Plate bottom is cuboid thin plate, and bottom is provided with rag iron spring.
4. according to claim 1 be used for tumour cell nucleic acid aptamers screening cartridge device, it is characterised in that the gas
Air filtering core is provided with the side air flue interface of road interface and liquid-transfering gun point.
5. according to claim 1 be used for tumour cell nucleic acid aptamers screening cartridge device, it is characterised in that the sky
The filter sizes of air filter film are less than aptamers.
6. according to claim 1 be used for tumour cell nucleic acid aptamers screening cartridge device, it is characterised in that described thin
Born of the same parents' culture dish adapter is semiclosed bulge, and bottom is provided with shaft.
7. according to claim 6 be used for tumour cell nucleic acid aptamers screening cartridge device, it is characterised in that described turn
Axis is seamlessly connected with bulge, the two is to be connected as a single entity, and shaft one end projects to the housing exterior of cartridge bodies, forms shaft
Prominent control point.
8. according to claim 1 be used for tumour cell nucleic acid aptamers screening cartridge device, it is characterised in that the examination
Agent storage reacting hole position includes the 9 holes position being successively set on cartridge reagent storage disk, and 9 holes position is respectively that library is high
Gentle discharge hole, library low-temp storage hole, with reference to liquid receiving hole, TE buffer solution receiving hole, waste liquid receiving hole, cleaning solution receiving hole,
Pancreatin digestive juice receiving hole, PCR reacting holes, aptamers transfer receiving hole.
9. according to claim 1 be used for tumour cell nucleic acid aptamers screening cartridge device, it is characterised in that the examination
Agent storage reacting hole position is circular or square, and bottom is distributed in hemispherical or taper ending closure, the orifice center point of hole position
On the circumference that the center of circle is overlapped with the turntable center of circle.
10. according to claim 1 be used for tumour cell nucleic acid aptamers screening cartridge device, it is characterised in that described
Library high temperature receiving hole 29 is detachable movable span position with aptamers transfer receiving hole 37, and the opening of hole position is closed with cork.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711472080.0A CN108004324A (en) | 2017-12-29 | 2017-12-29 | One kind is used for tumour cell nucleic acid aptamers screening circular card box device |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711472080.0A CN108004324A (en) | 2017-12-29 | 2017-12-29 | One kind is used for tumour cell nucleic acid aptamers screening circular card box device |
Publications (1)
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| CN108004324A true CN108004324A (en) | 2018-05-08 |
Family
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| CN201711472080.0A Pending CN108004324A (en) | 2017-12-29 | 2017-12-29 | One kind is used for tumour cell nucleic acid aptamers screening circular card box device |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108977344A (en) * | 2018-09-21 | 2018-12-11 | 西安佰奥莱博生物科技有限公司 | A kind of separating extraction device and separating and extracting process |
| CN110331075A (en) * | 2019-02-02 | 2019-10-15 | 上海思路迪医学检验所有限公司 | Closed sequencing library prepares cartridge |
| CN112608813A (en) * | 2020-12-21 | 2021-04-06 | 上海思路迪生物医学科技有限公司 | Multi-degree-of-freedom library preparation card box with external power source and method |
| CN114395476A (en) * | 2022-01-02 | 2022-04-26 | 李志清 | Pollution-free sample processing system before single cell sequencing and accurate processing system |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106353492A (en) * | 2016-10-13 | 2017-01-25 | 东南大学 | Cassette device for screening tumor cell aptamers |
| CN206114666U (en) * | 2016-10-13 | 2017-04-19 | 东南大学 | Be used for box -packed putting of tumor cells nucleic acid aptamer screening card |
-
2017
- 2017-12-29 CN CN201711472080.0A patent/CN108004324A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106353492A (en) * | 2016-10-13 | 2017-01-25 | 东南大学 | Cassette device for screening tumor cell aptamers |
| CN206114666U (en) * | 2016-10-13 | 2017-04-19 | 东南大学 | Be used for box -packed putting of tumor cells nucleic acid aptamer screening card |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108977344A (en) * | 2018-09-21 | 2018-12-11 | 西安佰奥莱博生物科技有限公司 | A kind of separating extraction device and separating and extracting process |
| CN110331075A (en) * | 2019-02-02 | 2019-10-15 | 上海思路迪医学检验所有限公司 | Closed sequencing library prepares cartridge |
| CN112608813A (en) * | 2020-12-21 | 2021-04-06 | 上海思路迪生物医学科技有限公司 | Multi-degree-of-freedom library preparation card box with external power source and method |
| CN112608813B (en) * | 2020-12-21 | 2022-07-01 | 上海思路迪生物医学科技有限公司 | Multi-degree-of-freedom library preparation card box with external power source and method |
| CN114395476A (en) * | 2022-01-02 | 2022-04-26 | 李志清 | Pollution-free sample processing system before single cell sequencing and accurate processing system |
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