CN107446802A - A kind of micro 384 hole DNA synthesizing and purifying plates - Google Patents
A kind of micro 384 hole DNA synthesizing and purifying plates Download PDFInfo
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- CN107446802A CN107446802A CN201710881882.0A CN201710881882A CN107446802A CN 107446802 A CN107446802 A CN 107446802A CN 201710881882 A CN201710881882 A CN 201710881882A CN 107446802 A CN107446802 A CN 107446802A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
Abstract
The invention discloses a kind of micro 384 hole DNA synthesizing and purifying plates, including synthetic vectors and 384 hole hollow plates, 384 orifice plate plate bodys are uniformly equally spaced 384 holes, each hole forms by the square bore portion of upper end and the circular bore portions of lower end through conical transition, assembling position is provided with 384 orifice bores, assembling position is the cube or cylinder of standard.The present invention solves the problem of micro denier DNA synthesis scales；Solve the problems, such as that the synthetic vectors volume of DNA synthesis is excessive；Solve the problems, such as that CPG dosages are excessive in carrier；Solve the problems, such as that reagent dosage is excessive in DNA building-up processes, saves synthetic agent dosage；The design of optimization solves the problems, such as 384 pore plate by sealing, compatible, reduce the mutation rate of building-up process, make product quality relatively reliable, solve the complicated processes that synthetic DNA product needs individually to purify simultaneously, the DNA products that can be directly washed on 384 orifice plates required for cutting us react for PCR, convenient and swift.
Description
Technical field
This new invention is related to minim DNA synthesis technical field, a kind of micro 384 hole DNA synthesizing and purifying plates.
Background technology
At present, DNA synthesizes post and often uses single-column pipe or 96 hole filters, and the specification of existing single-column pipe is more, and volume is big,
It is unfavorable for mass, scale and miniaturization operation, is synthesized for micro, ultramicron, high-volume DNA, single-column pipe is difficult to realize；
Existing 96 hole filters, mainly there are 1.5 and 2ml two kinds of specifications, while the synthetic work of 96 DNA samples can be completed, its
Synthesis for mass DNA, it is fast many compared with single tube, but in the sample size of processing, still can not meet micro, ultra micro
Measure DNA mass, scale is combined to.The shortcomings that existing single-column pipe and 96 hole filters, specifically it is also manifested by following several
Individual aspect：1）It is limited to synthesize sample size, once tests, simultaneously synthesizing 192 DNA samples of most multipotency；2）DNA synthesis scale all exists
More than 25nmol, can not meet micro ultramicron DNA synthesis, so needed for supporting DNA synthesis reagent dosage also very
Greatly, reagent consumption is not only increased, adds processing cost, and reagent treatment used is to human body and environmental toxic mostly
Harmful volatile corrosive liquids, dosage is more, and discharging of waste liquid is more, bigger to human body and environmental hazard；3）Automate journey
Degree is not high, inefficient, and operation is extremely inconvenient, can not realize automation, mass synthetic DNA sample well；4）Itself is produced
Product volume is with regard to big, and required matched reagent and filler matrix are also more, and the production cost of product in itself also remains high；5）Behaviour
Make complexity, after the synthesis of DNA products, it is also necessary to which follow-up cumbersome purification step, waste time and energy cost of idleness and resource；6）Function list
One, can not meet the needs of synthesizing and purifying integration.
However, with biotechnology, life science, synthetic biology, biological medicine, chemical analysis, food inspection and face
The fast development of bed diagnostics research to substantial amounts of DNA sample, it is necessary to carry out synthesizing and purifying, the instrument supporting to DNA synthesizing and purifyings
Device equipment and reagent consumptive material propose higher requirement, such as synthesis of high flux, ultramicron and detection, cost control, environmental protection, with
And usage amount of DNA synthesizing and purifying reagents etc. is reduced to greatest extent.
The content of the invention
It is an object of the invention to provide a kind of micro 384 hole DNA synthesizing and purifying plates, to solve to carry in above-mentioned background technology
The problem of going out.
To achieve the above object, the present invention provides following technical scheme：
A kind of micro 384 hole DNA synthesizing and purifying plates, including synthetic vectors and 384 hole hollow plates, 384 orifice plate plate bodys uniformly equidistantly divide
Cloth 384 holes, and each hole forms by the square bore portion of upper end and the circular bore portions of lower end through conical transition, 384 holes
At least one assembling position is provided with plate hole, assembling position is the cube or cylinder of standard.Inner chamber gradient beyond assembling position subtracts
Small, synthetic vectors are divided into two kinds of cube and cylinder, can be filled in the cube assembling position and cylinder assembling position of 384 orifice plates, side
The shape synthetic vectors length of side≤4.0mm, thickness 0.1mm-7mm, cylindrical synthetic vectors diameter≤2.0mm, thickness 0.1mm-
7mm, carrier scale 0-10nmol.
As further scheme of the invention：Assembling position >=1 in each hole in 384 orifice plates, top assembling position are side
Body structure, bottom assembling position is cube or cylindrical structure.
As further scheme of the invention：Described square bore portion is provided with least one assembling position, assembles position
The inner chamber length of side≤4.0mm, the height for assembling position are 0.1mm-7mm, and assembling position is square or rectangular parallelepiped structure.
As further scheme of the invention：Described circular bore portions are provided with least one assembling position, the assembling position
Internal diameter（Diameter）≤ 3.5mm, the height for assembling position are 0.1mm-7mm, and the assembling position is cylindrical structure.
As further scheme of the invention：384 described hole hollow plates are 384 single-column pipes or 4 piece of 96 hole filters
It is made.
As further scheme of the invention：The external dimensions of 384 described hole hollow plates：A length of 120mm-130mm, it is wide
For 80mm-88mm, a height of 10mm-40mm.
As further scheme of the invention：384 described hole hollow plate bottom point mouths are divided into angle or flat mouth or point
Mouth structure.
As further scheme of the invention：The 384 hole hollow plate circumferential side frame width is 0.1-3mm.
As further scheme of the invention：Assemble empty using cone between position cube and assembling position cylindrical cavity
Chamber transition, cone cavity height is between 1-20mm.
As further scheme of the invention：The square synthetic vectors length of side≤4.0mm, thickness 0.1mm-7mm, cylinder
Shape synthetic vectors diameter≤2.0mm, thickness 0.1mm-7mm.
As further scheme of the invention：Between carrier scale 0-10nmol.
As further scheme of the invention：Sintetics can be used for PCR reactions after directly cleaning cutting, without
Additional purification process.
Compared with prior art, the beneficial effects of the invention are as follows：
1st, solves the problem of micro denier DNA synthesis scales
The micro 384 hole DNA synthesizing and purifying plates of the present invention, it is to aim at minigene synthesis（0.1-3nmol）And design, synthesis rule
Mould minimum as little as 0.01nmol, and commercially available DNA synthesis post synthesis scales are general all in 25nmol or more.
2nd, solve the problems, such as that the CPG carrier bulks of DNA synthesis are excessive
Presently commercially available CPG carriers, diameter and thickness are substantially all in more than 2mm, and a kind of micro 384 hole DNA of the present invention is closed
Into purifying plate, diameter and thickness are all within 2mm, even as low as 1.2mm, for the DNA synthesis carriers of " minimum " in the world.
3rd, solve the problems, such as that CPG dosages are excessive in carrier
For the carrier of gene chemical synthesis, its CPG content just disclosure satisfy that the requirement of gene chemical synthesis between 1-50ug, but at present
Commercially available synthetic vectors content mostly in more than 1mg, some even in more than 10mg, the DNA products being synthesized considerably beyond
Actual demand, causes the serious wasting of resources, adds research and production cost, is unfavorable for building economizing type society
Meeting, and it is of the invention, and CPG contents can be with as little as 1ug in carrier.
4th, solve the problems, such as that reagent dosage is excessive in DNA building-up processes, saves synthetic agent dosage
Within a kind of micro 384 hole DNA synthesizing and purifying plates of the present invention, diameter and thickness as little as 1.2mm, reagent volume used
Minimum as little as 0.4ul, and the reagent volume required for traditional synthetic vectors at least needs more than 5ul, the synthetic agent of saving
At least more than 10 times of dosage.The technology not only reduces reagent dosage, saves production cost, has saved resource, has also protected
Environment.
5th, the design of optimization solves the problems, such as 384 pore plate by sealing, compatible, reduces the mutation rate of building-up process,
Make product quality relatively reliable.
Brief description of the drawings
Fig. 1 is the overall structure diagram of micro 384 hole DNA synthesizing and purifying plates.
Fig. 2 is part A enlarged diagram in Fig. 1 of micro 384 hole DNA synthesizing and purifying plates.
Fig. 3 is another overall schematic of micro 384 hole DNA synthesizing and purifying plates.
Fig. 4 is the schematic rear view of micro 384 hole DNA synthesizing and purifying plates.
Fig. 5 is the front schematic view of micro 384 hole DNA synthesizing and purifying plates.
Fig. 6 is the schematic cross-section of micro 384 hole DNA synthesizing and purifying plates.
Fig. 7 is another schematic cross-section of micro 384 hole DNA synthesizing and purifying plates.
Fig. 8 is the side sectional view of synthetic vectors in micro 384 hole DNA synthesizing and purifying plates
Fig. 9 is the top view of synthetic vectors in micro 384 hole DNA synthesizing and purifying plates.
Embodiment
The technical scheme of this patent is described in more detail with reference to embodiment.
Refer to Fig. 1-7, a kind of micro 384 hole DNA synthesizing and purifying plates, including synthetic vectors and 384 hole hollow plates, 384 holes
Plate plate body is uniformly equally spaced 384 holes, and each hole is by the square bore portion of upper end and the circular bore portions of lower end through circle
Cone transition is formed, and at least one assembling position is provided with 384 orifice bores, and assembling position is the cube or cylinder of standard.Assemble position
Inner chamber gradient in addition reduces；Synthetic vectors are divided into two kinds of cube and cylinder, can be filled in 384 orifice plates cube assembling position and
Cylinder assembles position.The square synthetic vectors length of side≤4.0mm, thickness 0.1mm-7mm, cylindrical synthetic vectors diameter≤
2.0mm, thickness 0.1mm-7mm, carrier scale 0-10nmol.
Further：The 384 hole hollow plate circumferential side frame width is 0.1-3mm.
Further：The 384 hole long 122.0mm of hollow plate, wide 81.0mm, high 18.0mm, plate surrounding frame width 3.0mm.
Further：The quantity for assembling position is 1, and the internal diameter for assembling position is 0.1mm-1.2mm, and the height for assembling position is
0.1mm-1.2mm, 384 hole hollow plate long 122.0mm, wide 81.0mm, high 18.0mm.
Further：Cone cavity transition is used between assembling position cylindrical cavity, cone cavity height is in 1-20mm
Between.
Embodiment 1
1st, the design of 384 holes synthesis hollow plate
Hollow plate long 122.0mm, wide 81.0mm, high 18.0mm, plate surrounding frame width 3.0mm, plate body are uniformly equally spaced 384
Hole, top square opening internal diameter length of side 3.59mm, gradually close up to length of side 3.5mm, 3.5mm and assembled under position to 3.5mm assemblings position top
Portion is gradually closed up from square opening and is transitioned into circular port, and until 0.9mm assembling position, 0.9mm assembling position bottom is gradually closed up extremely
0.6mm, bottom point mouth point is using angle or flat mouth or sharp-crested design.
2nd, the preparation of synthetic vectors（Calculated by preparing 10000）
1）Raw material：CPG carrying capacity is 20umol/g, and every 0.1nmol synthetic vectors amount containing CPG is about 5ug；UHMW-PE powder
Molecular weight be 3,000,000.
2）Raw material volume calculates：Every diameter 0.9mm thickness 1.0mm synthetic vectors volume is 0.63585ul, is pressed
10000 proportionings, cumulative volume is 6.3585ml ≈ 6.4ml.
3）Synthetic vectors raw materials for sintering configures：Weigh 50mg CPG and be put into 10ml graduated cylinders, be slowly added UHMW-PE powder and arrive
6.4ml scales, overturn and mix for several times, then with UHMW-PE polishings to 6.4ml positions；It is standby that mixing is overturned repeatedly.
4）Die-filling tool：In the aluminium-making mould in the hole containing 10000 diameter 0.9mm height 1.0mm, uniformly addition mixing
Good CPG and UHMW-PE blended powders, are shaken flat with vibration gauge, cover upper cover plate, are locked.
5）Sintering：200 DEG C, 23min is sintered, water cooling, takes out synthetic vectors.
3rd, assemble
Diameter 0.9mm thickness 1.0mm synthetic vectors are assembled in 384 holes synthesis hollow plate using full-automatic filling equipment
0.9mm assembles position.
This plate can be used as the hole DNA synthesizing and purifyings plates of 0.1nmol 384 to use.
Embodiment 2
1st, the design of 384 holes synthesis hollow plate
Hollow plate long 122.0mm, wide 81.0mm, high 17.22mm, plate surrounding frame width 3.0mm, plate body are uniformly equally spaced 384
Individual hole, top square opening internal diameter length of side 3.59mm, gradually closed up to length of side 3.5mm, 3.5mm assemblings position to 3.5mm assemblings position top
Bottom is gradually closed up from square opening and is transitioned into circular port, is cone cavity until 1.2mm assemblings position.Cone cavity lower end
For diameter 1.2mm, height 1.5mm cylindrical cavity assembling position, bottom is closed up to 0.8mm, ultimately forms a diameter of 0.8mm,
Highly bezel designs are used for 14mm cylindrical cavity to bottom, bottom point mouth point.
2nd, the preparation of synthetic vectors（Calculated by preparing 5000）
1）Raw material：CPG carrying capacity is 25umol/g, and every 1nmol synthetic vectors amount containing CPG is about 40ug；UHMW-PE powder
Molecular weight is 3,000,000.
2）Raw material volume calculates：Every diameter 1.2mm thickness 1.0mm synthetic vectors volume is 1.1304ul, by 5000
Proportioning, cumulative volume is 5.625ml ≈ 5.6ml.
3）Synthetic vectors raw materials for sintering configures：Weigh 200mg CPG and be put into 10ml graduated cylinders, be slowly added UHMW-PE powder
To 5.6ml scales, overturn and mix for several times, then with UHMW-PE polishings to 5.6ml positions；It is standby that mixing is overturned repeatedly.
4）Die-filling tool：In the aluminium-making mould in the hole containing 5000 diameter 1.2mm height 1.0mm, uniformly addition mixing
Good CPG and UHMW-PE blended powders, are shaken flat with vibration gauge, cover upper cover plate, are locked.
5）Sintering：190 DEG C, 30min is sintered, water cooling, takes out synthetic vectors.
3rd, assemble
Diameter 1.2mm thickness 1.0mm synthetic vectors are assembled in 384 holes synthesis hollow plate using full-automatic filling equipment
1.2mm assembles position.
This plate can be used as the hole DNA synthesizing and purifyings plates of 1nmol 384 to use.
Embodiment 3
1st, the design of 384 holes synthesis hollow plate
Hollow plate long 122.0mm, wide 81.0mm, high 17.22mm, plate surrounding frame width 3.0mm, plate body are uniformly equally spaced 384
Individual hole, top square opening internal diameter length of side 3.59mm, gradually closed up to length of side 3.5mm, 3.5mm assemblings position to 3.5mm assemblings position top
Bottom is gradually closed up from square opening and is transitioned into circular port, is cone cavity until 1.2mm assemblings position.Cone cavity lower end
For diameter 1.2mm, height 1.5mm cylindrical cavity assembling position, bottom is closed up to 0.8mm, ultimately forms a diameter of 0.8mm,
Highly bezel designs are used for 14mm cylindrical cavity to bottom, bottom point mouth point.
2nd, the preparation of synthetic vectors（Calculated by preparing 5000）
1）Raw material：CPG carrying capacity is 60umol/g, and every 3nmol synthetic vectors amount containing CPG is about 50ug；UHMW-PE powder
Molecular weight is 3,000,000.
2）Raw material volume calculates：Every diameter 1.2mm thickness 1.0mm synthetic vectors volume is 1.1304ul, by 5000
Proportioning, cumulative volume is 5.625ml ≈ 5.6ml.
3 synthetic vectors raw materials for sintering configure：Weigh 250mg CPG and be put into 10ml graduated cylinders, be slowly added UHMW-PE powder and arrive
5.6ml scales, overturn and mix for several times, then with UHMW-PE polishings to 5.6ml positions；It is standby that mixing is overturned repeatedly.
4）Die-filling tool：In the aluminium-making mould in the hole containing 5000 diameter 1.2mm height 1.0mm, uniformly addition mixing
Good CPG and UHMW-PE blended powders, are shaken flat with vibration gauge, cover upper cover plate, are locked.
5）Sintering：190 DEG C, 35min is sintered, water cooling, takes out synthetic vectors.
3rd, assemble
Diameter 1.2mm thickness 1.0mm synthetic vectors are assembled in 384 holes synthesis hollow plate using full-automatic filling equipment
1.2mm assembles position.
This plate can be used as the hole DNA synthesizing and purifyings plates of 3nmol 384 to use.
In the description of the invention, it is necessary to explanation, unless otherwise indicated, " multiple " be meant that two or two with
On；The orientation of the instruction such as term " on ", " under ", "left", "right", " interior ", " outer ", " front end ", " rear end ", " head ", " afterbody "
Or position relationship is based on orientation shown in the drawings or position relationship, is for only for ease of the description present invention and simplifies description, and
It is not instruction or implies that signified position there must be specific orientation, with specific azimuth configuration and operation, therefore can not manages
Solve as limitation of the present invention.In addition, term " first ", " second ", " the 3rd " etc. be only used for describe purpose, and it is not intended that
Instruction implies relative importance.
In the description of the invention, it is also necessary to explanation, unless otherwise clearly defined and limited, term " transition ",
" connected ", " connection " should be interpreted broadly, for example, can be to be fixedly connected between square opening and circular port in 384 orifice plates, also may be used
To be to be detachably connected, or it is integrally connected；384 holes can be mechanically connected；Can be joined directly together, can also be in
Between medium be indirectly connected.For the ordinary skill in the art, visual concrete condition understands above-mentioned term in the present invention
In concrete meaning.
The better embodiment of this patent is explained in detail above, but this patent is not limited to above-mentioned embodiment party
Formula, can also be on the premise of this patent objective not be departed from one skilled in the relevant art's possessed knowledge
Make a variety of changes or change, and these changes or modification, the essence of appropriate technical solution is departed from each reality of the present invention
Apply the spirit and scope of a technical scheme.
Claims (10)
1. a kind of micro 384 hole DNA synthesizing and purifying plates, it is characterised in that including synthetic vectors and 384 hole hollow plates, 384 orifice plate plates
Body is uniformly equally spaced 384 holes, and each hole is by the square bore portion of upper end and the circular bore portions of lower end through circular cone mistake
Cross and form, at least one assembling position is provided with 384 orifice bores, assembling position is the cube or cylinder of standard；Assemble beyond position
Inner chamber gradient reduce, synthetic vectors are divided into two kinds of cube and cylinder, can be filled in cube assembling position and the cylinder of 384 orifice plates
Body assembles position, and the square synthetic vectors length of side≤4.0mm, thickness 0.1mm-7mm, cylindrical synthetic vectors diameter≤2.0mm are thick
Spend 0.1mm-7mm, carrier scale 0-10nmol.
2. micro 384 hole DNA synthesizing and purifying plates as claimed in claim 1, it is characterised in that the dress in each hole in 384 orifice plates
Coordination >=1, top assembling position are cube structure, and bottom assembling position is cube or cylindrical structure.
3. micro 384 hole DNA synthesizing and purifying plates as claimed in claim 1, it is characterised in that described square bore portion is provided with
At least one assembling position, the inner chamber length of side≤4.0mm of position is assembled, the height for assembling position is 0.1mm-7mm, and assembling position is square
Or rectangular parallelepiped structure；Described circular bore portions are provided with least one assembling position, the internal diameter of the assembling position（Diameter）≤ 3.5mm,
The height for assembling position is 0.1mm-7mm, and the assembling position is cylindrical structure.
4. micro 384 hole DNA synthesizing and purifying plates as claimed in claim 1, it is characterised in that 384 described hole hollow plates are 384
Individual single-column pipe or 4 piece of 96 hole filters are made；The external dimensions of 384 hole hollow plates：A length of 120mm-130mm, a width of 80mm-
88mm, a height of 10mm-40mm.
5. micro 384 hole DNA synthesizing and purifying plates as claimed in claim 1, it is characterised in that 384 described hole hollow plate bottoms
Sharp mouth is divided into angle or flat mouth or sharp-crested structure.
6. micro 384 hole DNA synthesizing and purifying plates according to claim 1, it is characterised in that the 384 hole hollow plate surrounding
Border width is 0.1-3mm.
7. micro 384 hole DNA synthesizing and purifying plates according to claim 1, it is characterised in that assembling position cube and assembling position
Cone cavity transition is used between cylindrical cavity, cone cavity height is between 1-20mm.
8. micro 384 hole DNA synthesizing and purifying plates as claimed in claim 1, it is characterised in that the square synthetic vectors length of side≤
4.0mm, thickness 0.1mm-7mm, cylindrical synthetic vectors diameter≤2.0mm, thickness 0.1mm-7mm.
9. micro 384 hole DNA synthesizing and purifying plates as claimed in claim 1, it is characterised in that carrier scale 0-10nmol it
Between.
10. micro 384 hole DNA synthesizing and purifying plates as claimed in claim 1, it is characterised in that sintetics directly cleaning is cut
PCR reactions can be used for after cutting, without additional purification process.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710881882.0A CN107446802A (en) | 2017-09-26 | 2017-09-26 | A kind of micro 384 hole DNA synthesizing and purifying plates |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710881882.0A CN107446802A (en) | 2017-09-26 | 2017-09-26 | A kind of micro 384 hole DNA synthesizing and purifying plates |
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| CN107446802A true CN107446802A (en) | 2017-12-08 |
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ID=60497497
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| CN201710881882.0A Pending CN107446802A (en) | 2017-09-26 | 2017-09-26 | A kind of micro 384 hole DNA synthesizing and purifying plates |
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| CN110724623A (en) * | 2019-09-10 | 2020-01-24 | 深圳市百迈生命科学有限公司 | 1536 well solid phase synthesis and extraction plate |
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| CN111036166A (en) * | 2019-12-30 | 2020-04-21 | 江苏东玄基因科技有限公司 | Synthesis device and preparation method of spherical solid phase reaction carrier |
| CN116143447A (en) * | 2022-12-22 | 2023-05-23 | 河北迪纳兴科生物科技有限公司 | DNA synthesis column and preparation method thereof |
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