CN204022828U - The device of extracting nucleic acid molecule - Google Patents
The device of extracting nucleic acid molecule Download PDFInfo
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- CN204022828U CN204022828U CN201420410932.9U CN201420410932U CN204022828U CN 204022828 U CN204022828 U CN 204022828U CN 201420410932 U CN201420410932 U CN 201420410932U CN 204022828 U CN204022828 U CN 204022828U
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- nucleic acid
- filter cotton
- acid molecule
- tubing string
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Abstract
The utility model relates to a kind of device of extracting nucleic acid molecule, and it is for the device by cell or tissue purifying one nucleic acid molecule, comprising: a tubing string and a filter cotton.Wherein this filter cotton has the function of filtering the solidifying flocks producing in this non-chromosome nucleic acid molecule process of purifying, this device utilizes micropipet to see through filter cotton and filters the solidifying flocks producing in purification of nucleic acid molecules process, and draw the clear liquor contain target nucleic acid molecules, simplify several centrifugal process required in traditional purifying, to reduce the costs such as time of required cost and manpower.
Description
Technical field
The utility model is about a kind of device, particularly a kind of device that uses the extracting nucleic acid material in tubing string with filter cotton of nucleic acid substances extraction.
Background technology
After nineteen fifty-three, DNA double thigh spirane structure was untied, many resources constantly drop into the research about the each side such as structure, function and purposes of the nucleic acid molecule such as DNA and RNA, the particularly application in diagnosis and treatment disease aspect and genetic engineering about nucleic acid molecule.
And in the research of genetic engineering, with choosing, to grow specific objective DNA fragmentation be a kind of very common method as carrier to using the plastid of bacterium.Plastid forms the ability that ring-type is present in cell and has self-replacation conventionally.Develop at present many methods that can be used to extraction or purifying plastid DNA, comprised alkaline bleach liquor cleavage method, boiling method, cesium chloride method of purification and commercialization external member etc.Wherein alkaline bleach liquor cleavage method is a kind of higher and more easy method of efficiency, and cardinal principle, for utilizing alkaline matter to make cell endoplast and karyomit(e) sex change, makes DNA be split into sub-thread by bifilar solution, the acidic substance of then take again neutralization make DNA by sub-thread renaturation as bifilar.Yet in this renaturation reaction, small molecules plastid DNA can revert to rapidly base pairing mode and configuration originally in renaturation reaction, has compared with highly water-soluble; Yet the chromosomal DNA that molecular weight is larger cannot match the configuration that recovers original form a kind of solidifying cotton-shaped white precipitate with the sylvite in solution smoothly, thus can profit in such a way by target plastid DNA and chromosome segregation.
Yet, solidifying cotton-shaped still need separated with the settled solution that is dissolved with target plastid DNA of precipitation just can be completed through several centrifugation step, if can be by these centrifugal process simplifications in the process of automatization, there is great help for reduction time and human cost.Moreover, the extraction process of the nucleic acid molecule of other kinds, as while extracting the nucleic acid molecule of vegetable cell or tissue, the inclusion of cell or tissue lysate can be divided into water-soluble and water-insoluble, and nucleic acid molecule is dissolved in water-soluble portion, also after non-water-soluble impurity first must being got rid of with centrifugation, water miscible nucleic acid molecule is being isolated, this centrifugal process also needs to simplify to save time and human cost.
Utility model content
In order to address the above problem, the purpose of this utility model is to provide a kind of device of extracting nucleic acid molecule, and it is simple in structure, easy to operate, and energy simplification process, to reduce costs, raises the efficiency.
For achieving the above object, the utility model discloses a kind of device of extracting nucleic acid molecule, it is characterized in that comprising: a tubing string and a filter cotton, this filter cotton is clogged in tubing string.
Wherein, this nucleic acid molecule is plastid, DNA or RNA.
Wherein, this filter cotton is positioned at this tubing string bottom.
Wherein, the pore size of this filter cotton is between 0.2 to 20 micron.
Wherein, the pore size of this filter cotton is between 0.5 to 10 micron.
Wherein, this filter cotton is embedded in the tube wall of this tubing string bottom.
According to above-mentioned device of the present utility model, its advantage producing compared to prior art is as follows: device of the present utility model utilizes micropipet to see through filter cotton and filters the solidifying flocks producing in purification of nucleic acid molecules process, and draw the clear liquor contain target nucleic acid molecules, simplify several centrifugal process required in traditional purifying, to reduce the costs such as time of required cost and manpower.
Accompanying drawing explanation
Fig. 1: the schematic diagram that shows the device of the utility model extracting nucleic acid material.
Embodiment
Term " one " or " a kind of ", when being used in conjunction in claim or specification sheets with " comprising ", may represent one herein.But also meet " one or more " or " at least one ".
Please refer to Fig. 1, it shows the schematic diagram of the device of the utility model extracting nucleic acid material.
The purpose of this utility model is mainly to provide a kind of device 100 of extracting nucleic acid molecule, comprising: a tubing string 40 and a filter cotton 30, this filter cotton 30 is clogged in tubing string 40.Wherein, the better nucleic acid molecule that can be used for extracting in cell of device 100 of the present utility model, be somebody's turn to do " cell " term, represent a kind of prokaryote, bacterial cell for example, or a kind of eukaryotic cells, for example yeast cell, vegetable cell, mammalian cell (as human cell or mouse cell), it has been known unrestrictedly its kind.Moreover, device of the present utility model also can be used for extracting the nucleic acid molecule in a tissue, term in the utility model " tissue ", for the cell framework between cell and organ, by many plesiomorphic cells and intercellular substance, formed such as epithelium, muscle tissue, reticular tissue or nervous tissue etc.
Above-mentioned term " nucleic acid molecule " or " nucleic acid ", be to take Nucleotide as the molecule that unit was formed, and comprises DNA, RNA etc.On the one hand, device of the present utility model is better for extracting or the nucleic acid molecule of purifying tissue; Herein term " extraction or purifying tissue in nucleic acid molecule ", the nucleic acid molecule that means this extraction does not comprise in fact cell debris or intercellular substance, therefore, nucleic acid molecule herein can comprise karyomit(e) molecule and non-chromosome molecule.Herein alleged " target nucleic acid molecules " can because of extraction or the object of purifying different, and represent different nucleic acid molecule, as being while extracting plastid in bacterium when object, this target nucleic acid molecules represents achromosomal nucleic acid molecule; And when object when extracting karyomit(e) nucleic acid molecule in animal or plant is organized, target nucleic acid molecules represents the karyomit(e) nucleic acid molecule of this animal or plant.Therefore, in technical field, have and conventionally know the knowledgeable under the utility model, can be because of different extractions or purifying object, and understand the connotation of target nucleic acid molecules herein.
On the other hand, device of the present utility model is preferably for extracting or purifying cells karyomit(e) nucleic acid molecule in addition, as plastid DNA, RNA etc.Term " extraction or purifying cells in a non-chromosome nucleic acid molecule " herein, the non-chromosome nucleic acid molecule that means this extraction does not comprise in fact cell debris and chromosomal DNA.
Above-mentioned term " filter cotton 30 ", is positioned at this tubing string 40 bottoms, and it act as and filters solidifying cotton-shaped throw out, and in other words, it can be by the particle of specific size or impurity filtering in a liquid solution.Term " solidifying flocculent precipitate " herein, if in cell when extraction or purifying plastid DNA, means to be consisted of the water-fast material of chromosomal DNA, cytolemma, protein and other cell debriss etc. that forms solidifying wadding with acidic substance; If when extraction or purify DNA or RNA, organize lysate can form " fibery precipitate " in tissue, mean to be formed by cytolemma, protein, intercellular substance and other cell debriss etc. water-fast material.
The pore size of above-mentioned filter cotton 30, between 0.2 to 20 micron, is preferably between 0.5 to 10 micron, to avoid aperture excessive and solid-state solidifying flocculent substance is mixed with the settled solution of target via filter cotton 30 holes.
Above-mentioned filter cotton 30, it is embedded in the tube wall of these tubing string 40 bottoms substantially completely.Wherein " completely chimeric substantially " refer to the tube wall tabling of 90% above area and tubing string 40 bottoms, more preferably more than 95%, as 95%, 96%, 97%, 98%, 99% or in fact 100% chimeric." tubing string 40 " in herein can be the tubing string 40 using in known laboratory, and it comprises a lid 10 (if lid nonessential also can omit) and a body 20, and this body 20 is better but be not limited to cylindrical.For making filter cotton 30 be embedded in substantially the tube wall of these tubing string 40 bottoms completely, filter cotton 30 is preferably the shape identical with tubing string 40, as is a cylindrical filter cotton 30; But consider the elasticity of filter cotton 30 own, can use a square filter cotton 30, wherein this square length of side is slightly larger than the diameter of circular tubing string 40, while square filter cotton 30 being filled in to this tubing string bottom, because the elasticity of filter cotton 30 makes so, first compressed and filled in behind tubing string bottom, filter cotton 30 100% tube wall that is embedded in tubing string 40 bottoms in fact after the bottom space of tubing string 40 expands.The thickness of this filter cotton 30 is also unrestricted, as long as can effectively filter solid-state solidifying flocculent substance wherein; Preferably thickness is 1/6th height of tubing string 40 length, or 1/5th height.
Above-mentioned filter cotton 30, wherein material is selected the group that free Mierocrystalline cellulose, glass fibre and polymkeric substance form, but does not limit its kind, and this material is known.
The nucleic acid molecule purification process that device of the present utility model carries out
Further illustrate the nucleic acid molecule purification process that utilizes device 100 of the present utility model to carry out, comprise: (a) cell or is organized to cracking, to form the suspension with target nucleic acid molecules; (b) make the non-target nucleic acid molecules impurity in this suspension form a solidifying flocculent precipitate; And (c) through these filter cotton 30 these solidifying flocculent precipitates of filtration of this device 100, the clear liquid state material of only drawing containing this target nucleic acid molecules carries out nucleic acid purification step.
Wherein step (a) the method for cell or tissue cracking can be comprised physical method as homogenized, ultrasound, French cell crushing instrument; Chemical process is as chemical bacteriolyze (cultivating as used 0.2M NaOH and 1%SDS mixed solution) etc.; Or ferment method is as used N,O-Diacetylmuramidase etc.During above-mentioned cell or tissue cracking, can add RNase A.Conventionally when cell or tissue cracking, first cell or tissue is suspended in a suspension, the formula of this suspension has been known and can be by buying on the market, as glucose, Tris-HCl and EDTA mixed solution.
The main purpose of above-mentioned cell or tissue cleavage method is to make cytolemma/wall to be damaged, and intracytoplasmic material is disengaged and/or destroys intercellular substance, forms a lysate/molten cytosol that comprises the materials such as cytoplasm protein and plastid.
Then, in step (b), make the non-target nucleic acid molecules impurity in this suspension form a fibrous or solidifying flocculent precipitate, this method that forms solidifying flocculent precipitate is also known, as use acidic substance as Potassium ethanoate or sodium-acetate etc., make chromosomal DNA form solidifying flocculent precipitate.Non-target nucleic acid molecules impurity in suspension forms fibery precipitate or solidifying flocculent precipitate is while extracting target nucleic acid molecules difference, and form different throw outs.In an embodiment, if the plastid DNA in wish extraction bacterial cell can add acidic substance to make chromosomal DNA and acid form solidifying flocculent precipitate in suspension; In another embodiment, if want, in tissue, extract chromosomal DNA, will organize after homogeneous, chromosomal DNA is water-soluble, has the fibery precipitate that non-target nucleic acid molecules impurity forms as intercellular substance, reticular tissue fragment etc. in this suspension.
According to the step (c) of the above, can utilize known method that the clear liquid state material containing this target nucleic acid molecules is carried out to nucleic acid purification step, its purification step is also known, as uses known tubing string method of purification purification of target nucleic acid; Wherein, better micropipet or the syringe with filtering membrane that utilize directly drawn filtered this clear liquid state material in this device.Wherein, should be with in the micropipet or syringe of filtering membrane, this filtering membrane has the function of adsorption of DNA or RNA, can make the clear liquor after filter cotton filters pass through after the filtering membrane of micropipet, target dna molecule in clear liquor or RNA molecule are adsorbed on this filtering membrane, and then easily that the target dna molecule in clear liquor or RNA molecule is separated with other unwanted liquid or impurity.(should be known with the micropipet of filtering membrane, can be with reference to TaiWan, China patent announcement M477925.)
According to the step of the above, in a better enforcement aspect, when wherein this micropipet is inserted this device 100, carry out exhaust, when this micropipet props up this filter cotton 30, start to draw this clear liquid state material.The object of wherein carrying out steps of exhausting is to avoid micropipet to be drawn to because of carelessness fibrous or solidifying cotton-shaped solid matter in the middle of propping up the front process of this filter cotton 30, owing to comprising water-fast cytolemma, protein and other cell debriss etc. in fibrous or solidifying flocculent substance, if when drawing it and mixing with target settled solution, can cause the Efficiency Decreasing of purification of target DNA or RNA.Moreover, when this micropipet props up this filter cotton 30, be preferably and directly will there is elastic filter cotton 30 and be pressed onto near pipe the end (approximately distance pipe the end 0.01～0.1cm), make this micropipet equal the filter tip function that front end has filter cotton 30, and can guarantee that the liquid that micropipet sucks is all the settled solutions after filtering via filter cotton 30, but this micropipet can not be depressed into filter cotton 30 the pipe end completely and this micropipet props up the pipe end, otherwise cannot draw solution.
In addition, step of the present utility model (a), (b) and (c) all can in device 100 of the present utility model, carry out, or step (a) can be prior to carrying out in an Eppendorf tube (eppendorf), and then suspension being drawn to device 100 of the present utility model, and the step (b) of chatting after carrying out in device 100 and (c).Or, can be after step (b) have carried out, then all liquid is drawn to device 100 of the present utility model.
In sum, known of the present utility model being characterised in that utilized purification of nucleic acid device 100, comprise a tubing string 40 and a filter cotton 30, utilize this device 100 to filter the fibrous or solidifying flocks producing in nucleic acid molecule purge process, reduce the costs such as required time of centrifugal process and manpower.
Following examples should not be considered as exceedingly limiting the utility model.Under the utility model, in technical field, have and conventionally know that the knowledgeable can modify and change embodiment discussed in this article in the situation that not deviating from spirit of the present utility model or category, and still belong to scope of the present utility model.
Embodiment 1 to 3
Use the plastid extraction cover group of device 100 collocation QIAGEN Plasmid Min Kit (Catalog no.12123) of the present utility model.First after the intestinal bacteria liquid of the 2ml in Eppendorf tube is centrifugal, remove supernatant liquor, then add the P1 damping fluid of 300 μ l, with pipettor, aspirate up and down and make cell Eddy diffusion.Suspension is transferred in device 100 of the present utility model, adds the P2 damping fluid of 300 μ l, with pipettor, aspirate up and down and make lysis, then, then add in 300 P3 damping fluid suspension and produce solidifying floss.(it is automated installation to use micropipet with filtering membrane, can auto-pumping) exhaust limit, limit is close toward the filter cotton 30 of the bottom of device 100, wait propping up this filter cotton 30 and during from tubing string 40 bottom 0.01cm, stop exhaust, change into absorption, the solution with plastid DNA is drawn to this micropipet, to a clean centrifuge tube, and further aspirate and make plastid DNA be adsorbed in the filtering membrane on micropipet up and down, and waste liquid is got rid of, then aspirate respectively 750 μ l scavenging solution (BufferQC) twice, with cleaning and filtering film, micropipet is aspirated and pushed away and drain into few 15 times, make residual in the alcohol volatilization in micropipet.Finally, 200 μ l are rushed to extract (Buffer QF) via the absorption of this micropipet, and standing 3 minutes, then, that discharges this molten plastid DNA rushes extract to the Eppendorf tube of 1.5ml.Three repetitions are carried out in each experiment, are respectively embodiment 1 to 3.
Comparative example 1 to 3
Use the plastid extraction cover group of the label identical with embodiment, first after the intestinal bacteria liquid of the 2ml in Eppendorf tube is centrifugal, remove supernatant liquor, then the P1 damping fluid that adds 300 μ l, with pipettor, aspirate up and down and make cell Eddy diffusion, then add the P2 damping fluid of 300 μ l, rapidly after reversion repeatedly, add again after the P3 damping fluid reversion repeatedly of 300 μ l, in suspension, produce solidifying floss.Then, after centrifugal 10 minutes, draw supernatant liquor, and use the plastid DNA in QIAGEN Plasmid Min Kit (Catalog no.12123) extraction cover group extraction supernatant liquor.Three repetitions are carried out in each experiment, are respectively comparative example 1 to 3.
It in following table 1, is the result test that embodiment and comparative example extract plastid DNA, visible use device of the present utility model carries out plastid extraction, the plastid DNA concentration that it extracts and purity and prior art difference are little, but can simplify preposition centrifugation step numerous and diverse in prior art, and can carry out automatic decimation, and have, save time and the better effect of manpower.
Table 1
| Sample | O.D.230 | O.D.260 | O.D.280 | 260/280 | 260/230 |
| Embodiment 1 | 0.278 | 0.565 | 0.314 | 1.80 | 2.04 |
| Embodiment 2 | 0.276 | 0.554 | 0.310 | 1.79 | 2.01 |
| Embodiment 3 | 0.267 | 0.521 | 0.293 | 1.78 | 1.95 |
| Comparative example 1 | 0.327 | 0.624 | 0.347 | 1.80 | 1.91 |
| Comparative example 2 | 0.285 | 0.560 | 0.313 | 1.79 | 1.97 |
| 0.285 | 0.560 | 0.313 | 1.79 | 1.97 | 1.92 |
Claims (6)
1. a device for extracting nucleic acid molecule, is characterized in that comprising: a tubing string and a filter cotton, this filter cotton is clogged in tubing string.
2. device as claimed in claim 1, is characterized in that, this nucleic acid molecule is plastid, DNA or RNA.
3. device as claimed in claim 1 or 2, is characterized in that, this filter cotton is positioned at this tubing string bottom.
4. device as claimed in claim 3, is characterized in that, the pore size of this filter cotton is between 0.2 to 20 micron.
5. device as claimed in claim 4, is characterized in that, the pore size of this filter cotton is between 0.5 to 10 micron.
6. device as claimed in claim 3, is characterized in that, this filter cotton is embedded in the tube wall of this tubing string bottom.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201420410932.9U CN204022828U (en) | 2014-07-24 | 2014-07-24 | The device of extracting nucleic acid molecule |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201420410932.9U CN204022828U (en) | 2014-07-24 | 2014-07-24 | The device of extracting nucleic acid molecule |
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| CN204022828U true CN204022828U (en) | 2014-12-17 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019001228A1 (en) * | 2017-06-30 | 2019-01-03 | 开启基因股份有限公司 | Fixed tube of nucleic acid extraction component, and nucleic acid extraction component |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019001228A1 (en) * | 2017-06-30 | 2019-01-03 | 开启基因股份有限公司 | Fixed tube of nucleic acid extraction component, and nucleic acid extraction component |
| US11365406B2 (en) | 2017-06-30 | 2022-06-21 | Catchgene Co., Ltd. | Fixed tube of nucleic acid extraction component, and nucleic acid extraction component |
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| C14 | Grant of patent or utility model | ||
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| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20141217 Termination date: 20210724 |