CN104130941A - Centrifugal sleeve tube structure used for separating and extracting DNA - Google Patents
Centrifugal sleeve tube structure used for separating and extracting DNA Download PDFInfo
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- CN104130941A CN104130941A CN201310159320.7A CN201310159320A CN104130941A CN 104130941 A CN104130941 A CN 104130941A CN 201310159320 A CN201310159320 A CN 201310159320A CN 104130941 A CN104130941 A CN 104130941A
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- 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
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1017—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by filtration, e.g. using filters, frits, membranes
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Abstract
A centrifugal sleeve tube structure used for separating and extracting DNA belongs to the field of equipment used for separation and purification of nucleic acid DNA or RNA. The structure includes a centrifugal tube. A connecting ribbon is formed on an upper part of the centrifugal tube. A plug cover is connected to an end of the connecting ribbon. An inner sleeve tube is disposed in a centrifugal tube chamber. A liquid discharge groove is formed on a bottom wall of an inner sleeve tube chamber and one side, which is opposite to the inner sleeve tube chamber, of the bottom wall is extended to form an inner sleeve connecting port. An impurity-filtering mechanism includes a transition sleeve cylinder and a coil. A transition sleeve cylinder matching joint is formed on a central position of the bottom of a transition sleeve cylinder chamber, wherein the center of the joint forms a liquid discharge hole. The coil is disposed in the transition sleeve cylinder chamber with one end being matched with the inner sleeve connecting port and the other end being matched with the transition sleeve cylinder matching joint. A coil chamber of the coil is filled with filtering materials. By means of the structure, a secondary pipetting process is unnecessary so that a water-soluble DNA template is free from losing; and an impurity-removing effect is excellent and a PRC amplification operation is ensured. The structure is convenient, quick to operate, is labor-saving, is good in the impurity-removing effect, is simple and is easy to manufacture.
Description
Technical field
The invention belongs to nucleic acid DNA or RNA separation and purification equipment technical field, be specifically related to the centrifuge shield structure that a kind of separation and Extraction DNA uses.
Background technology
Though separation and purification extract DNA be medical treatment, criminal case investigation and to all kinds of accidents as fields such as traffic accident victim's identification, archaeologies or all there is positive meaning in animals and plants inspection and quarantine field, the detection method of DNA is main but be not limited to following several: phenol-chloroform extraction process, chelex-100 method; Paramagnetic particle method; Organic solvent method; Salting-out process and silica bead method, etc.No matter but adopt which kind of method, its common feature or claim common requirement roughly can be summarized as following two aspects: the one, as far as possible farthest improve the output of DNA or RNA and purity and PCR is reacted to inhibition and be reduced to minimum degree; The 2nd, simple and effective, cost is low and it is little to pollute.
In aforesaid DNA detection method, phenol-chloroform extraction process was once the method for generally using, but because the DNA of phenol-chloroform extraction process complex operation, extraction exists the protein contamination that is difficult to avoid and because phenol and chloroform have the toxicity of harmful to human and be eliminated gradually.
Because silica bead method has efficiently, cost is low and be tending towards the strong point such as pollution-free, thereby be generally subject to the approval of industry, this method is first sample to be introduced in centrifuge tube, and in centrifuge tube, add TES(Pehanorm base ethyl sulfonic acid, be called for short biological buffer) and SLS(sodium laurylsulfonate salt); Reheat cracking and add adsorption liquid, then after high speed centrifugation, adopt pipettor to move pipe, namely the material in centrifuge tube moved in another centrifuge tube and add silica bead standing; Finally use successively rinsing liquid rinsing, ethanol rinsing and TC(elutriant) wash-out.But, silica bead method is extracted DNA and in actual operating process, is had following shortcoming: one, owing to existing, use pipettor negative pressure to move pipe ring joint, therefore in moving pipe process, the liquid that had previously been arranged in centrifuge tube can not fully be transferred to another centrifuge tube, thereby causes water miscible DNA profiling loss large (conventionally reaching more than 30%); Its two, owing to moving with pipettor in pipe process, the impurity in original centrifuge tube can not fully be removed, thus affect pcr amplification (polymerase chain reaction, English name is: Polymerase Chain Reaction); Its three, owing to existing, manually move pipe ring joint, therefore be still not enough to a certain extent embody time saving and energy saving and effect efficiently; Its four, the diluting effect to adsorption liquid due to TES and SLS, thus the concentration of adsorption liquid is significantly declined (70% left and right that is about original concentration), have a strong impact on the ability of silica bead adsorption of DNA; Its five, the double influence owing to being subject to aforementioned DNA profiling loss and TES and SLS to adsorption liquid dilution, significantly declines the concentration of DNA profiling (being only 50% left and right nearly of original concentration, even lower than 50%).
Fairly obvious, the shortcoming that is not limited to the aforementioned silica bead method extraction DNA exemplifying can ascribe moving due to pipe in operating process to, the problems referred to above more particularly, if abandoned by pipettor, the liquid rotating in previous centrifuge tube moved to another centrifuge tube in operating process, just can be readily solved so.For this reason, the applicant has carried out literature search, Chinese patent Granted publication CN201744365U recommends to have " filter-membrane centrifugal tube ", although this patent scheme can embody the technique effect described in its specification sheets the 0007th hurdle, and the description from its 0009 hurdle, in operating process without moving pipe, but because structure is lost the reasonable following drawback that exists: the one, impurity easily enters the bottom of centrifuge tube with body fluid from filter membrane (patent claims blend fiber millipore filtration), thereby the purity of water miscible DNA profiling is exerted an influence, the 2nd, due to the rubber ring of shelving for filter membrane be difficult to and the inwall of centrifuge tube between form desirable sealing, therefore the microgap of the same meeting of impurity between rubber ring and the inwall of centrifuge tube, enter the bottom of centrifuge tube, because must meeting with the tube chamber chamber wall of centrifuge tube, the rubber ring of this patent instruction can plug requirement, so, if reach ultimate attainment sealing effectiveness in order to meet between the tube chamber chamber wall of rubber ring and centrifuge tube, in follow-up operation steps, be difficult to so, by the link on rubber ring, rubber ring is taken out to centrifuge tube together with pressure ring and filter membrane, otherwise, if for indemnifying party just takes out rubber ring, certainly will affect so the sealing effectiveness between the tube chamber chamber wall of rubber ring and centrifuge tube, the 3rd, owing to only objectively cannot embodying with one piece of filter membrane the good effect filtering out impurities, thereby affect pcr amplification.
Granted publication CN202610231U provides " multilayer nucleic acid absorption centrifuge shield ", because this patent scheme is contrary with aforementioned patent to the extraction principle of DNA, what adopt is absorption method, although in operating process without moving pipe, but the loss of the template of DNA is difficult to control, specifically can be referring to specification sheets the 0025th hurdle of this patent.
In view of above-mentioned prior art, be necessary to be improved, technical scheme described below produces under this background.
Summary of the invention
Task of the present invention is to provide a kind of and contributes to abandon secondary and move pipe and use and avoid the loss of water miscible DNA profiling, be conducive to fully remove impurity and use the amplification that ensures PCR, be of value to avoiding relying on and manually move pipe and use and embody quick and laborsaving in operating process, have and be convenient to prevent TES, SLS exerts an influence to the concentration of adsorption liquid and uses and promote the adsorptive power of silica bead to DNA, have be good at fully retaining original DNA profiling and use significantly improve the recall rate of low copy biological material and one-piece construction simple and use the centrifuge shield structure that the convenient separation and Extraction DNA manufacturing uses.
Task of the present invention completes like this, and the centrifuge shield structure that a kind of separation and Extraction DNA uses comprises and forms the centrifuge tube there is centrifugal tube chamber, on the top of this centrifuge tube, forms and has a connecting band, at the end of this connecting band, is connected with a gag, one inner sleeve, this inner sleeve is placed in the centrifugal tube chamber of described centrifuge tube, and form and have a drain tank in the edge of the diapire of the internal canula lumen of this inner sleeve, at diapire, back to the edge of a side of internal canula lumen, be extended with an inner sleeve interface, the inner sleeve hub cavity of this inner sleeve interface communicates with described drain tank, one for introducing described centrifugal intraluminal Lv Za mechanism by the impurity filtering of the DNA sample of being drawn by described inner sleeve interface and by aqueous solution DNA profiling after filtering out impurities, Gai Lvza mechanism comprises a transition sleeve and a coil pipe, in the bottom in the transition sleeve chamber of this transition sleeve and be positioned at middle position and form and have a transition sleeve connector, the central authorities of this transition sleeve connector form a leakage opening, this leakage opening communicates with described centrifugal tube chamber, coil pipe is arranged in transition sleeve chamber, and one end of this coil pipe and described inner sleeve interface connect, and the other end and described filter sleeve connector connect, in the coil pipe chamber of coil pipe, be provided with filter material.
In a specific embodiment of the present invention, in the centrifugal tube chamber of described centrifuge tube and be positioned on the wall body of bottom of centrifugal tube chamber and form and have a narrow contracting step, described filter sleeve is found a place on narrow contracting step.
In another specific embodiment of the present invention, on the outer wall of described filter sleeve and be positioned at bottom and form and to have one group of support foot, this group support foot to be shelved on described narrow contracting step with interval state.
In another specific embodiment of the present invention, in the lower end of described inner sleeve, form and have an external diameter to be less than the narrow contracting Connection Block of the external diameter of inner sleeve, and form on the top of the wall body in the transition sleeve chamber of described transition sleeve, have a mating cavity, this mating cavity matches with narrow contracting Connection Block.
In another specific embodiment of the present invention, described Transition Materials is protein filter membrane.
Technical scheme provided by the invention is with respect to one of technique effect of prior art, owing to being provided with inner sleeve in the centrifugal tube chamber of centrifuge tube, and Lv Za mechanism is placed in centrifugal tube chamber in company with inner sleeve, therefore the water-soluble DNA profiling after centrifugal and the filter of You Lvza mechanism are mixed can enter in the centrifugal tube chamber of centrifuge tube, and the impurity You Lvza mechanism filtering such as protein in DNA sample, therefore when subsequent operations, need only inner sleeve is withdrawn to centrifuge tube together with Lv Za mechanism, without carrying out secondary, move pipe, thereby can avoid the loss of water miscible DNA profiling, two, because Jiang Lvza mechanism is configured on inner sleeve, so impurity-eliminating effect is desirable and be conducive to ensure the amplification of PCR, three, therefore owing to moving pipe without secondary, can fully demonstrate easy to operate, quick and labour-saving strong point, four, due to good impurity removing effect, thereby can prevent that TES, SLS from exerting an influence to adsorption liquid concentration and ensureing the adsorptive power of silica bead to DNA, can fully retain original DNA profiling again and use the recall rate that significantly improves low copy biological material, five, because Lv Za mechanism only consists of transition sleeve and coil pipe, thereby simple in structure, be easy to make.
Accompanying drawing explanation
Fig. 1 is embodiments of the invention structure iron.
Fig. 2 is the sectional view of Fig. 1.
Embodiment
For the auditor that the makes Patent Office especially public can be expressly understood technical spirit of the present invention and beneficial effect more, applicant elaborates the mode with embodiment below, but to the description of embodiment, be not all the restriction to the present invention program, any according to the present invention design, done only for pro forma but not substantial equivalent transformation all should be considered as technical scheme category of the present invention.
Embodiment 1:
Refer to Fig. 1 and Fig. 2, provided a centrifuge tube 1, this centrifuge tube 1 is preferably used transparent plastics molded and shaped, on the chamber wall of the centrifugal tube chamber 11 of this centrifuge tube 1 and the portion on the lower side that is positioned at the short transverse of centrifugal tube chamber 11 form and have a narrow contracting step 13.Shown in figure, the narrow contracting of the diameter of the bottom of centrifugal tube chamber 11 and form and have a reservoir compartment 111, the water miscible DNA profiling of being stored after removal of impurities by this reservoir compartment 111, to drop into silica bead and adsorb in follow-up operation steps.At the position of the accent of the centrifugal tube chamber 11 of centrifuge tube 1, form and have a connecting band 12, and the end of this connecting band 12 is connected with a gag 121.
As preferred scheme, also can be on the outer wall of centrifuge tube 1 and form a centrifuge tube mounting flange limit 14 in the surrounding of the accent corresponding to centrifugal tube chamber 11, by this centrifuge tube mounting flange limit 14, ensure that centrifuge tube 1 matches with centrifugal device (not illustrating in the drawings because belonging to known technology).
Provided an inner sleeve 2, the same transparent plastics that preferably uses of this inner sleeve 2 is molded and shaped, on the diapire 211 of these inner sleeve 2 internal canula lumen 21 and around the edge of the circumferential direction of diapire 211, form and have a surperficial drain tank 2111 being recessed in diapire 211, at diapire 211, back to a side of internal canula lumen 21, towards the edge of the side in discrete core barrel chamber 11, be extended with an inner sleeve interface 2112, the inner sleeve hub cavity 21121 of this inner sleeve interface 2112 communicates with drain tank 2111, liquid in internal canula lumen 21 enters inner sleeve hub cavity 21121 through drain tank 2111.In the bottom of inner sleeve 2, form and have the narrow contracting Connection Block 22 that an external diameter is less than the external diameter of inner sleeve 2.
Please continue to refer to Fig. 1 and Fig. 2, provide Liao Yilvza mechanism 3, Gai Lvza mechanism 3 comprises a filter sleeve 31 and a coil pipe 32, on the diapire in transition sleeve 31 transition sleeve chambeies 311 and be positioned at middle position and form and to have a transition sleeve connector 3111 that is raised in bottom wall surface, the central authorities of this transition sleeve connector 3111 form leakage opening 31111, and leakage opening 31111 communicates with centrifugal tube chamber 11.On the top in the filter sleeve chamber 311 of filter sleeve 31, form and have a mating cavity 3112, this mating cavity 3112 and aforesaid narrow contracting Connection Block 22 coordinate (Fig. 2 shows) on the outer wall of filter sleeve 31 and are positioned at bottom and form and have one group of support foot 312, this group support foot 312 to be bearing on aforesaid narrow contracting step 13 with interval state.Peg graft and coordinate with inner sleeve interface 2112 in one end of coil pipe 32, the other end is pegged graft and coordinated with filter sleeve connector 3111, is provided with the filter material 321 of being filled the post of by protein filter membrane in the coil pipe chamber of coil pipe 32.
From diagram, when inner sleeve 2 be contained in the centrifugal tube chamber 11 of centrifuge tube 1 interior after, clip locating flange 23 on inner sleeve 2 reliably contacts with the chamber wall of the centrifugal tube chamber 11 of centrifuge tube 1, and after DNA sample introducing internal canula lumen 21 is interior, by aforesaid gag 121 lids, is fitted over the position of the inner sleeve mouth of pipe of internal canula lumen 21.
Preferably, at the top of inner sleeve 2, on the outer wall in the position of the accent corresponding to internal canula lumen 11, to extending out, be provided with an inner sleeve flange 24, this inner sleeve flange 24 is friendly with aforesaid gag 121 to be coordinated.
Applicant sketches use of the present invention, the first interior introducing of internal canula lumen 21 DNA sample, TES and SLS to inner sleeve 2, carry out again heating pyrolyze and add adsorption liquid, then inner sleeve 2 is inserted in the centrifugal tube chamber 11 of centrifuge tube 1 together with Lv Za mechanism 3, and gag 121 cap seals, in the inner sleeve mouth of pipe position of the internal canula lumen 21 of inner sleeve 2, are then placed in centrifuge tube 1 on centrifugal device.Under the high-speed motion of centrifugal device, liquid in internal canula lumen 21 is through drain tank 2111 and inner sleeve hub cavity 21121, and the filter material in coil pipe 32 321 is that protein filtering membrane filters, through leakage opening 31111, enter aforesaid reservoir compartment 111, in this process, because impurity Jing Lvza mechanism 3 filters in DNA sample are assorted, the water-soluble NDA template therefore entering in reservoir compartment 111 is not only lost little, and impurity eliminated substantially, for follow-up silica bead absorption provides technical guarantee.Because impurity-eliminating effect is very good, so DNA profiling retains the recall rate that fully can improve low copy biological material.After centrifugal end, inner sleeve 2 is removed from centrifugal tube chamber 11 together with Lv Za mechanism 3, enter subsequent step and as the aforementioned silica bead is dropped into reservoir compartment 111 and adsorb etc.
In sum, technical scheme provided by the invention has objectively made up the shortcoming in prior art, has completed invention task, has cashed the technique effect described in the superincumbent technique effect of applicant hurdle.
Claims (5)
1. the centrifuge shield structure that separation and Extraction DNA uses, it is characterized in that comprising that formation has a centrifuge tube (1) of centrifugal tube chamber (11), on the top of this centrifuge tube (1), form and have a connecting band (12), at the end of this connecting band (12), be connected with a gag (121), one inner sleeve (2), this inner sleeve (2) is placed in the centrifugal tube chamber (11) of described centrifuge tube (1), and form and have a drain tank (2111) in the edge of the diapire (211) of the internal canula lumen (21) of this inner sleeve (2), at diapire (2111), back to the edge of a side of internal canula lumen (21), be extended with an inner sleeve interface (2112), the inner sleeve hub cavity (21121) of this inner sleeve interface (2112) communicates with described drain tank (2111), one for introducing the Lv Za mechanism (3) in described centrifugal tube chamber (11) by the impurity filtering of the DNA sample of being drawn by described inner sleeve interface (2112) and by the aqueous solution DNA profiling after filtering out impurities, Gai Lvza mechanism (3) comprises a transition sleeve (31) and a coil pipe (32), in the bottom in the transition sleeve chamber (311) of this transition sleeve (31) and be positioned at middle position and form and have a transition sleeve connector (3111), the central authorities of this transition sleeve connector (3111) form a leakage opening (31111), this leakage opening (31111) communicates with described centrifugal tube chamber (11), coil pipe (32) is arranged in transition sleeve chamber (311), and one end of this coil pipe (32) and described inner sleeve interface (2112) connect, and the other end and described filter sleeve connector (3111) connect, in the coil pipe chamber of coil pipe (32), be provided with filter material (321).
2. the centrifuge shield structure that separation and Extraction DNA according to claim 1 uses, it is characterized in that in the centrifugal tube chamber (11) of described centrifuge tube (1) and be positioned on the wall body of bottom of centrifugal tube chamber (11) forming and having a narrow contracting step (13), described filter sleeve (31) is found a place on narrow contracting step (13).
3. the centrifuge shield structure that separation and Extraction DNA according to claim 2 uses, it is characterized in that on the outer wall of described filter sleeve (31) and be positioned at bottom forming and having one group of support foot (312) with interval state, this group support foot (312) is shelved on described narrow contracting step (13).
4. the centrifuge shield structure that separation and Extraction DNA according to claim 1 uses, it is characterized in that forming and having an external diameter to be less than the narrow contracting Connection Block (22) of the external diameter of inner sleeve (2) in the lower end of described inner sleeve (2), and form on the top of the wall body in the transition sleeve chamber (311) of described transition sleeve (31), have a mating cavity (3112), this mating cavity (3112) matches with narrow contracting Connection Block (22).
5. the centrifuge shield structure that separation and Extraction DNA according to claim 1 uses, is characterized in that described Transition Materials (321) is protein filter membrane.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310159320.7A CN104130941B (en) | 2013-05-03 | 2013-05-03 | The centrifuge shield structure of separation and Extraction DNA |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310159320.7A CN104130941B (en) | 2013-05-03 | 2013-05-03 | The centrifuge shield structure of separation and Extraction DNA |
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| CN104130941A true CN104130941A (en) | 2014-11-05 |
| CN104130941B CN104130941B (en) | 2015-11-25 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105935514A (en) * | 2016-05-31 | 2016-09-14 | 无锡市疾病预防控制中心 | Solution transfer-free sample centrifuge filter |
| WO2021026832A1 (en) * | 2019-08-14 | 2021-02-18 | 王俊超 | Microburette upper lid, kit comprising the microburette upper lid and method thereof |
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2013
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| US20110137020A1 (en) * | 2004-03-18 | 2011-06-09 | Roche Molecular Systems, Inc. | Method and Device for Purifying Nucleic Acids |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105935514A (en) * | 2016-05-31 | 2016-09-14 | 无锡市疾病预防控制中心 | Solution transfer-free sample centrifuge filter |
| CN105935514B (en) * | 2016-05-31 | 2017-11-28 | 无锡市疾病预防控制中心 | A kind of sample centrifugal filter that need not shift solution |
| WO2021026832A1 (en) * | 2019-08-14 | 2021-02-18 | 王俊超 | Microburette upper lid, kit comprising the microburette upper lid and method thereof |
Also Published As
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|---|---|
| CN104130941B (en) | 2015-11-25 |
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Effective date of registration: 20151008 Address after: Changshou City ancient town Baimao Furong Village in Suzhou City, Jiangsu province 215532 Applicant after: CHANGSHU LEIHAO MEDICAL DEVICES CO., LTD. Address before: Changshou City ancient town Baimao Economic Development Zone of Suzhou city in Jiangsu province 215532 204 National Highway and Shanghai Yi section Applicant before: Changshu Kangbao Medical Appliance Factory |
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