CN203048936U - Nucleic acid purifying column system - Google Patents
Nucleic acid purifying column system Download PDFInfo
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- CN203048936U CN203048936U CN 201320067250 CN201320067250U CN203048936U CN 203048936 U CN203048936 U CN 203048936U CN 201320067250 CN201320067250 CN 201320067250 CN 201320067250 U CN201320067250 U CN 201320067250U CN 203048936 U CN203048936 U CN 203048936U
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- preparative column
- nucleic acid
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Abstract
The utility model provides a nucleic acid purifying column system which comprises two biological sample preparation columns which are independent to each other and can be overlapped together up and down; and each biological sample preparation column comprises a body, wherein the upper part of the body is provided with a sample feeding hole, the lower part of the body is provided with a sample outlet pipe nozzle, and a holding component is arranged at the upper part of the sample feeding hole in the biological sample preparation column; and the lower part of the upper biological sample preparation column is closely buckled in the holding component of the lower biological sample preparation column. According to the nucleic acid purifying column system, as the purification columns which can be overlapped to each other are adopted, the continuous operation of experiments and the conversion effect among different samples are met.
Description
Technical field
The utility model relates to biological sample and extracts the field, relates in particular to a kind of nucleic acid purification column system for nucleic acid extraction.
Background technology
In field of biology, often need filter and purifying sample.Especially in biology field, often need to extract the macromolecular substance such as DNA, RNA in the biological sample, and these macromolecular substance are carried out separation and purification.The separation and purification nucleic acid substances becomes the gordian technique of biology field expeditiously.Utilize preparative column from biological sample, to separate and extract nucleic acid and become biology field means commonly used.Common preparative column is the tubular structure of a circle, comprises the round tube cylinder, and an end of cylinder has sealing cover, and the other end is filled with porous sieve plate or film, has added the material that can adsorb biological substances such as nucleic acid at how empty sieve plate.Normal negative pressure method and the centrifuging of adopting extracted nucleic acid.When adopting the negative pressure method, at first preparative column is inserted on the socket of negative pressure device, needs the sample that separates with the dissolving of nucleic acid binding buffer liquid, as PCR product, enzyme cut, enzyme is marked or the product and being transferred in the preparative column of checking order, open and regulate negative pressure to the-25-30 inch of mercury, slowly siphon away solution in the pipe.Keep negative pressure, add after washings washs the residue in the pipe, preparative column is placed new centrifuge tube, central authorities add elutriant or deionized water at the preparative column film, and room temperature leaves standstill the centrifugal 1min wash-out of 12,000 * g nucleic acid behind the 1min.When adopting centrifuging, need the sample that separates with the dissolving of nucleic acid binding buffer liquid, as PCR product, enzyme cut, enzyme is marked or the product that checks order, and transfers to behind the mixing in the preparative column, and preparative column is placed centrifuge tube, the centrifugal 1min of 12,000 * g abandons filtrate.Preparative column is put the centrifuge tube of Hui Xin, add washings, the centrifugal 1min of 12,000 * g abandons filtrate.Again preparative column is placed clean centrifuge tube, central authorities add a certain amount of elutriant or deionized water at the preparative column film, and the centrifugal 1min wash-out of 12,000 * g obtained nucleic acid after room temperature left standstill 1min.Be subjected to the influence of this body structure of preparative column in the existing nucleic acid extraction technology, every through a wash-out or centrifugal, preparative column all will place new centrifuge tube again, therefore the step of converting complexity between the different samples.The existing preparation post is to be used in combination mutually each other, therefore can not satisfy the continuity operation of nucleic acid extraction.
The utility model content
For overcoming the deficiencies in the prior art, the purpose of this utility model is to provide the nucleic acid purification column system, at least comprise that two separately can be gone up the next biological sample preparative column that is superimposed again, described biological sample preparative column comprises body, upper part of body has the application of sample mouth, the bottom of body has the sample of going out ozzle, and the application of sample mouth top of biological sample preparative column is provided with a holding member, and the preparative column bottom that wherein occupy the top closely is fastened in the preparative column holding member that occupy the below.
Preferably, it is identical with the external diameter of preparative column bottom above occuping to occupy the internal diameter of preparative column holding member of below.
Preferably, the periphery that goes out the sample ozzle that occupy the preparative column of top also comprises fixed frame, and described fixed frame inserts in the preparative column holding member that occupy the below, and the external diameter of this fixed frame is identical with the internal diameter of the holding member of the preparative column that occupy the below.
Preferably, preparative column goes out the sample ozzle and comprises exit end, and described exit end is exposed to fixed outer frame.
Preferably, the junction of preparative column holding member and body also comprises bearing surface.
Preferably, have filtering membrane or pellosil in the body of preparative column, or both combinations.
The beneficial effects of the utility model are: nucleic acid purification column system described in the utility model comprises that two separately can be gone up the next biological sample preparative column that is superimposed again.Top at preparative column has holding member, and the inside diameter of holding member is designed to identically with the lower outer diameter of same type preparative column, and perhaps the external diameter than the fixed frame of its big dissimilar preparative column is identical.When needs carried out the continuity operation, the bottom of one of them preparative column or fixed frame just can be fastened in the holding member of another preparative column closely, had realized the mode with a plurality of preparative column stacks.More excellent mode is that preparative column also comprises a bearing surface and since bearing surface to insert wherein another preparative column bottom or the effect of keeping out of preparative column fixed frame, further guaranteed two steadiness that preparative column fastens.Thereby realize between the preparative column of the same type or the mutual superposition between the dissimilar preparative column, satisfied the continuity operational requirement of experiment.Mutual superposition between the preparative column can reduce the step of centrifuge tube decon, biological sample is extracted and the continuity of operation of purifying stronger.Operation steps in the middle of reducing can reduce the product of leaching process to environment and operator's pollution and harm, also can avoid the operation of too much step can increase the probability that extracts sample contamination.The usage quantity of centrifuge tube also can significantly reduce in addition, saves cost.
Description of drawings
Fig. 1 is first kind of biological sample preparative column structural representation.
Fig. 2 is first kind of nucleic acid purification column system structural representation.
Fig. 3 is the partial enlarged drawing at Fig. 2 A place.
Fig. 4 is second kind of biological sample preparative column structural representation.
Fig. 5 is second kind of nucleic acid purification column system structural representation.
The partial enlarged drawing at Fig. 6 Fig. 5 B place.
Fig. 7 is biological sample preparative column vertical view, comprises inner diaphragm structure synoptic diagram.
Embodiment
Below in conjunction with concrete accompanying drawing the utility model is described in detail.These specific embodiments only are limited the enumerating under the utility model spirit, do not get rid of one of ordinary skill in the art prior art and the utility model in conjunction with and other specific embodiments of producing.
Biological sample preparative column 1 as shown in Figure 1 comprises the body 2 of hollow, and the top of described body 2 has application of sample mouth 3, and the bottom of body has the sample of going out ozzle 4, is provided with a holding member 5 on the top of application of sample mouth.Holding member is used for holding another preparative column and inserts wherein, and can block insertion another preparative column wherein tightly, thereby two preparative columns can be superimposed use.The inside diameter of holding member 5 can be according to the difference of inserting preparative column external diameter wherein and difference.If the preparative column of mutual superposition is same kind, then the inside diameter of holding member 5 is identical with the external diameter of preparative column body.If the preparative column of mutual superposition is same kind, but the body of this preparative column is up big and down small or up-small and down-big cone, and then the holding member of preparative column and this preparative column overlapping portion are the preparative column bottom external diameter is identical.If the preparative column of mutual superposition is dissimilar, then holding member 5 inside diameters are identical with the lower outer diameter of the preparative column that inserts another different model wherein.Holding member 5 also comprises bearing surface 6 with the junction of body 2 in the preferred scheme, and bearing surface 6 can be the structure of a level and holding member 5 and body interconnected.Bearing surface also can be that the ramp structure with certain angle interconnects holding member 5 and body.The inclined-plane gradient of bearing surface can be identical with the inclined-plane gradient of body bottom 7.
Preparative column also comprises sealing cover 8 in one embodiment, and sealing cover 8 is connected with body 2 by connecting arm 9.In a preferred scheme, the bottom of preparative column and go out between the sample ozzle 4 to comprise dividing plate 10.Dividing plate 10 structures as shown in Figure 7 are the reticulated structure of porous.
Be the nucleic acid purification column system as shown in Figure 2, comprise the biological sample preparative column mutual superposition of the same kind that two sizes are identical together.In this nucleic acid purification column system, the internal diameter of the holding member 5 of following preparative column 1 is identical with top preparative column 1 bottom outer diameter, make that occuping top preparative column can be inserted in the following preparative column, and tightly blocked by the holding member of following preparative column.Fig. 3 is the partial enlarged drawing of Fig. 2 A part, a part that occupy the body bottom 7 of top preparative column just in time leans against on the bearing surface 6 of the preparative column that occupy following, thereby further consolidated the fastening power between two preparative columns, two preparative columns are by firm being superimposed.
Of the present utility model another kind of biological sample preparative column 11 as shown in Figure 4 comprises the body 2 with cavity, the top of described body 2 has application of sample mouth 3, the bottom of body has the sample of going out ozzle 4, be provided with a holding member 5 on the top of application of sample mouth, the inside diameter of holding member 5 can be according to the difference of inserting preparative column external diameter wherein and difference.Holding member 5 also comprises bearing surface 6 with the junction of body 2, and bearing surface 6 can be the structure of a level or inclined-plane with constant slope.Also comprise fixed frame 12 in the periphery that goes out sample ozzle 4, a section of fixed frame 12 is connected on the bottom 7 of body.In a preferred scheme, the exit end that goes out the sample ozzle stretches out fixed frame 12, namely goes out sample ozzle 12 exit end and is exposed to fixed outer frame.Because the exit end at exit end has stretched out fixed outer frame, the frame that is not fixed surrounds, and therefore when biological sample extracted, the material that comes out from exit end can not be ejected on the fixed frame, has guaranteed the purity of extracting.Preparative column 11 also comprises sealing cover 8 in one embodiment, and sealing cover 8 is connected with body by connecting arm 9.In a preferred scheme, the bottom of preparative column 11 and go out between the sample ozzle 4 to comprise dividing plate 10.
Be the another kind of nucleic acid purification column system of the utility model as shown in Figure 5, comprise that the biological sample preparative column of two types inequality is superimposed.In this nucleic acid purification column system, occuping following is small volume preparative column 1, occupy the top large volume preparative column 11 that is.Wherein to go out the fixed frame outer wall diameter of sample ozzle periphery identical with inserting large volume preparative column wherein for the inner diameter of the holding member 5 of small volume preparative column, make that occuping top large volume preparative column 11 can be inserted in the small volume preparative column that occupy following, and lived by the holding member of small volume preparative column fastening tightly.Fig. 6 is the partial enlarged drawing of Fig. 5 A part, in preferred scheme, occuping the part that top large volume preparative column 11 goes out sample ozzle fixed frame 12 just in time is resisted against on the bearing surface 6 of the small volume preparative column 1 that occupy following, owing to the support of bearing surface, further guaranteed the stability of two preparative column mutual superposition.
The described nucleic acid purification column system of utility model can be used for extracting nucleic acid, as DNA, RNA, and biological substances such as protein, the impurity in the time of can being used for removing the biological sample pre-treatment etc.
When this biological sample extraction column be used for to extract nucleic acid, at preparative column body interior described in the utility model pack into nucleic acid adsorbing porous membrane, for example pellosil, glass fibre membrane, silica nano material, Graphene, titanium dioxide crystal film etc.These material tolerance acid-base can specific absorption nucleic acid, and the other biological material is not adsorbed substantially, can ensure DNA or RNA in the recovery sample farthest, removes other impurity simultaneously.Be 5.0 to 6.5 at pH, contain under the hypersaline environment of Guanidinium hydrochloride of 4M that pellosil is adsorption of DNA, RNA optionally.When being in the high pH environment of less salt following time, thereby the DNA, the RNA that are adsorbed on this pellosil can discharge the purpose that reaches purifying.The aperture of these porous-films is 0.1 micron to 5 microns, preferably 1 micron.The thickness of porous-film is 0.1 millimeter to 2 millimeters, preferred 1 millimeter thickness.
The material of preparative column described in the utility model is the polypropylene of medical grade, the strict control of production process, pollutions such as no DNA, Dnase, Rnase.
Nucleic acid purification column system of the present utility model can be used for the continuous negative pressure air-exhaust method and extracts nucleic acid.
Embodiment 1 high purity plasmid DNA is extracted in a large number
1. material and reagent
The nucleic acid purification column system is to be in the same place with the large volume preparative column mutual superposition that diameter is suitable for the centrifuge tube of 50mL by the small volume preparative column that diameter is suitable for the centrifuge tube of 15mL;
Pellosil: energy specific adsorption nucleic acid;
Filtering membrane: this filtering membrane can be combined with plasmid DNA generation specificity, and its aperture can make plasmid DNA pass through smoothly, and materials such as albumen, thalline can not pass through this filtering membrane;
Centrifuge tube.
Solution I: bacterial suspension comprises 2mmol/L KH
2PO
4, 10mmol/L Na
2HPO
4, 137mmol/L NaCl, 2.7mmol/L KCl, 1%Tween20, pH6.0-8.0 behind the adding RNase A, mixes 4 ℃ of storages;
Solution II: bacterial lysate comprises 0.1-2.5mol/L NaOH, the airtight storage of 0.05%-1%SDS room temperature;
Solution III: neutralizer comprises 0.1-0.5mol/L (NH
4)
2HPO
4.NH
4H
2PO
4, 0.5-3mol/L KCl, pH4.5;
Solution IV: washings comprises 200mmol/L Tris, 2mol/L EDTA, 1.5mol/LNaCl, 3mol/L GuHCl, the airtight storage of room temperature;
Solution V: the liquid that desalts comprises the 50-100% dehydrated alcohol;
Elutriant: 2.5mmol/L Tris-HCl, pH8.5, the airtight storage of room temperature;
RNase?A。
2. step
1) get the high copy number plasmid bacterium liquid of 30ml overnight incubation in the LB substratum, or the low copy plasmid/Cosmid bacterium liquid of 100ml incubated overnight (if use rich medium, bacteria liquid is long-pending should to reduce by half or still less), 〉=3, the centrifugal 8min of 000 * g abandons supernatant.Centrifuge tube was inverted on the paper handkerchief several minutes, eliminates supernatant.
2) add solution I (bacterial suspension has added RNase A) the suspension bacterial precipitation of 4.5ml, suspending needs evenly should not leave little bacterium piece.
3) add 4.5ml solution II (bacterial lysate), gentleness also spins upside down fully to mix for 6-8 time and makes the abundant cracking of thalline, until forming bright solution; This step should not surpass 5min.The solution III (neutralizer) that adds 4.5ml4 ℃ of precooling in the clear solution, gentle and spin upside down 10 times fully and mix, until the aggegation piece that forms consolidation; Room temperature is placed 5min.
4) as shown in Figure 5, large volume biological sample preparative column 11 is installed on the small volume biological sample preparative column 1, small volume preparative column 1 is inserted on the socket of negative pressure device.Gained solution in the step 3 is slowly added in the large volume preparative column, open and regulate negative pressure to the-25-30 inch of mercury, slowly siphon away solution in the pipe.
5) treat that solution in the step 4 blots after, the large volume preparative column is pulled out from the small volume preparative column, the large volume preparative column is because cleaved thalline relic is held back down in the existence of filtering membrane, and allows target substance such as nucleic acid smoothly by entering in the small volume preparative column.Continue to keep negative pressure, in the small volume preparative column, add 5ml solution IV (washings), exhaust solution in the small volume preparative column.
6) in the small volume preparative column, add 5ml solution V (liquid desalts), exhaust solution in the pipe.
7) the small volume preparative column in the step 6 is placed clean 15ml centrifuge tube, add 0.3ml solution V (liquid desalts), the centrifugal 5min of 8,000 * g
8) the small volume preparative column is placed another clean 15ml centrifuge tube, add 0.3ml elutriant or deionized water.Room temperature leaves standstill 1min.The centrifugal 2min of 8,000 * g collects plasmid DNA.
3. detect
Detect with agarose gel electrophoresis and ultraviolet spectrophotometry, the concentration and the purity that reclaim the gained plasmid DNA meet the requirements.
Method in the nucleic acid purification column system described in this embodiment 1 and extraction plasmid DNA thereof.Because the large volume preparative column in this nucleic acid purification column system comprises filtering membrane, this filtering membrane can be held back down cleaved thalline relic, and allows target substance such as nucleic acid smoothly by entering in the small volume preparative column.The existence of large volume preparative column has replaced removing with centrifugation method the step of thalline relic.Owing to large volume preparative column in this nucleic acid purification column system links to each other with the small volume preparative column, make target products such as removing thalline relic and purifying nucleic acid realize the operate continuously slitless connection again.Therefore having significantly reduced operation steps has improved efficient, has realized automatization.
Claims (6)
1. nucleic acid purification column system, at least comprise that two separately can be gone up the next biological sample preparative column that is superimposed again, described biological sample preparative column comprises body, upper part of body has the application of sample mouth, the bottom of body has the sample of going out ozzle, it is characterized in that the application of sample mouth top of biological sample preparative column is provided with a holding member, the preparative column bottom that wherein occupy the top closely is fastened in the preparative column holding member that occupy the below.
2. nucleic acid purification column system according to claim 1 is characterized in that, the internal diameter of the preparative column holding member below occuping is identical with the external diameter of the preparative column bottom that occupy the top.
3. nucleic acid purification column system according to claim 1, it is characterized in that, the periphery that goes out the sample ozzle that occupy the preparative column of top also comprises fixed frame, described fixed frame inserts in the preparative column holding member that occupy the below, and the external diameter of this fixed frame is identical with the internal diameter of the holding member of the preparative column that occupy the below.
4. nucleic acid purification column system according to claim 3 is characterized in that, preparative column goes out the sample ozzle and comprises exit end, and described exit end is exposed to fixed outer frame.
5. nucleic acid purification column system according to claim 1 is characterized in that, the junction of preparative column holding member and body also comprises bearing surface.
6. nucleic acid purification column system according to claim 1 is characterized in that, has filtering membrane or pellosil in the body of preparative column, or both combinations.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201320067250 CN203048936U (en) | 2013-02-05 | 2013-02-05 | Nucleic acid purifying column system |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201320067250 CN203048936U (en) | 2013-02-05 | 2013-02-05 | Nucleic acid purifying column system |
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| Publication Number | Publication Date |
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| CN203048936U true CN203048936U (en) | 2013-07-10 |
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| CN 201320067250 Expired - Lifetime CN203048936U (en) | 2013-02-05 | 2013-02-05 | Nucleic acid purifying column system |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103087903A (en) * | 2013-02-05 | 2013-05-08 | 杭州百迈生物技术有限公司 | Nucleic acid purifying column system and its application in nucleic acid extraction |
| CN103343088A (en) * | 2013-07-16 | 2013-10-09 | 苏州市公安局 | Deoxyribonucleic acid (DNA) splitting and carrier removal device for batch biological samples |
| CN105413233A (en) * | 2015-12-10 | 2016-03-23 | 中国医学科学院生物医学工程研究所 | Micro-column separation device |
| CN105602942A (en) * | 2016-03-23 | 2016-05-25 | 王智勇 | Nucleic acid releaser and method for quickly extracting nucleic acids from dried blood spots |
-
2013
- 2013-02-05 CN CN 201320067250 patent/CN203048936U/en not_active Expired - Lifetime
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103087903A (en) * | 2013-02-05 | 2013-05-08 | 杭州百迈生物技术有限公司 | Nucleic acid purifying column system and its application in nucleic acid extraction |
| CN103343088A (en) * | 2013-07-16 | 2013-10-09 | 苏州市公安局 | Deoxyribonucleic acid (DNA) splitting and carrier removal device for batch biological samples |
| CN105413233A (en) * | 2015-12-10 | 2016-03-23 | 中国医学科学院生物医学工程研究所 | Micro-column separation device |
| CN105413233B (en) * | 2015-12-10 | 2017-07-04 | 中国医学科学院生物医学工程研究所 | A kind of Micro-Column Separation device |
| CN105602942A (en) * | 2016-03-23 | 2016-05-25 | 王智勇 | Nucleic acid releaser and method for quickly extracting nucleic acids from dried blood spots |
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
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| CP03 | Change of name, title or address |
Address after: Hangzhou City, Zhejiang province 311228 Jiangdong Industrial Zone Linjiang high tech Zone weft five road No. 3688 building four layer 2 Branch Park Patentee after: HANGZHOU KBM LIFE SCIENCES CO.,LTD. Address before: 310007, A, East 628, Zhejiang Science Park, 525 Xixi Road, Hangzhou, Zhejiang, Xihu District Patentee before: HANGZHOU KBM LIFE SCIENCES CO.,LTD. |
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| CX01 | Expiry of patent term |
Granted publication date: 20130710 |