CN108456678A - A kind of method of rapid extraction genomic DNA - Google Patents
A kind of method of rapid extraction genomic DNA Download PDFInfo
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- CN108456678A CN108456678A CN201810737847.6A CN201810737847A CN108456678A CN 108456678 A CN108456678 A CN 108456678A CN 201810737847 A CN201810737847 A CN 201810737847A CN 108456678 A CN108456678 A CN 108456678A
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- C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
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Abstract
The invention discloses a kind of methods for capableing of rapid extraction genomic DNA, and the method includes using following molecular sieve pillars：The molecular sieve column attached bag includes shaft and is arranged is used to hold the pallet of the holder and closed bottom of DNA seperation films in shaft bottom；The DNA seperation films free-standing is placed on holder, and there are gap, the method and step using above-mentioned molecular sieve pillar rapid extraction genomic DNA includes for the DNA seperation films and shaft：Disrupted sample cell obtains the extract mixed liquor of DNA；And the step of using above-mentioned molecular sieve pillar more convenient efficient acquisitions genomic DNA, the method for extracting nucleic acid of molecular sieve pillar progress using the present invention are easy to operate, complicated instrument and equipment is not needed, at low cost, DNA output is high, it is high-quality, there is good industrial applications foreground.
Description
Technical field
The present invention relates to separation, preparation or the technical fields for purifying DNA, and in particular to a kind of rapid extraction genomic DNA
Method.
Background technology
There are many extracting method about genomic DNA, substantially there is following several types：1) precipitation method；2) pellosil is miniature
Column method；3) silica gel column chromatography method；4) paramagnetic particle method.Theirs is common that processing procedure of the early period to sample, will with lysate
Clasmatosis, core DNA are released, and (purpose of protease k processing is for protein by the protease k degradation overwhelming majority
Extracted DNA purity is further increased, to the sample that purity requirement is not stringent, this step can be omitted).In different extractions
In method, subsequent method divergence is larger.
1) precipitation method isopropanol or other organic reagents, and corresponding high salt component are added in sample so that DNA parents
Water-bound changes, and is precipitated at flocculence, by centrifugation DNA, redissolves in TE or other buffer solutions.
2) pellosil micro-column method, lysate sample are added to pellosil micro-column, and DNA is specifically adsorbed in pellosil
On, by centrifugation or vacuumizing method, other substances for removing lysate and not adsorbing other are further eluted using cleaning solution
Impurity, be eventually adding eluent, allow DNA and silica gel UF membrane, collect the DNA of purifying.
3) silica matrix is filled in chromatographic column by silica gel column chromatography method, allows sample dissociation liquid by pillar, and DNA is special
Property be adsorbed on silica gel, other impurity are further eluted with cleaning solution, are finally detached by DNA and pillar with eluent, are received
Collect the DNA of purifying；
4) paramagnetic particle method compares with others several method, and paramagnetic particle method occurs than later, and the major technique of paramagnetic particle method is special
Point is to combine silica gel and magnetic-particle, forms silica gel magnetic bead (the inside is magnetic bead, and outside is silica gel).The silica gel of outer layer can be with
DNA in sample dissociation liquid is specifically combined under hydrophobic conditions, and the magnetic bead of inner layer can be generated in magnet or electromagnetic force
Magnetic field in move, in fixed regional ensemble, and other non-DNA dirt solutions without absorption/attachment separate, so as to
Easily to carry out the substance that is not bound with of cleaning, DNA eluents are finally used, the hydrophobicity of DNA is made to change and silica gel
Separation, collects the DNA of purifying.Paramagnetic particle method can readily realize automation mechanized operation.
Above four kinds of methods cut both ways, and first is higher with the yield of the DNA product of the third method, but operate multiple
Miscellaneous, cost is higher, is suitble to do the processing of a small amount of sample in R＆D units, and the operating cost of second method is high, complex steps,
The yield of nucleic acid DNA is not high, but reliable in quality, relatively laboratory is suitble to use, fourth method can be in automatic nucleic acid extraction
It is applied on instrument, can realize the nucleic acid extraction of mass, the disadvantage is that DNA output is low, it is of high cost.It can it would therefore be highly desirable to develop one kind
The method of rapid extraction genomic DNA and its matching used novel molecular sieve pillar, so that nucleic acid extraction technology is met with
Lower requirement：1) be easy large-scale production, 2) may be implemented mass production, 3) can obtain high yield high-quality genomic DNA,
4) operating procedure is simple and the operating time is short and 5) at low cost.
In the prior art, the genome DNA extracting reagent kit (product identification 51104) that classical Qiagen methods use
Production cost is higher, in use in the presence of time-consuming, and needs the corollary equipments such as high speed centrifugal machine for minim or vacuum pump, deposits
Using not convenient enough problem.
Invention content
To solve the above-mentioned problems, the invention discloses a kind of method for capableing of rapid extraction genomic DNA, the methods
Include the steps that using following molecular sieve pillars：
The molecular sieve column attached bag includes shaft and DNA seperation films；The shaft bottom is provided with for holding DNA separation
The holder of film and the pallet of closed bottom；Wherein, the holder of the DNA seperation films, may be used X fonts, can also adopt
With other various frame shapes, as long as playing the role of holding DNA seperation films, for example, can also using the style of similar Fig. 2.
The DNA seperation films free-standing is placed on holder, and there are gaps for the DNA seperation films and shaft, described
DNA seperation films can isolate and fall off under slight outer force effect to shaft top orientation (one direction).From angle easy to produce
For degree, the diameter of DNA Separation membranes is slightly less than internal diameter 1mm of pillar or so.
Method and step using above-mentioned molecular sieve pillar rapid extraction genomic DNA includes：
1. then the isopropanol of 1/3-1/2 volumes is added in disrupted sample cell, the extract for being uniformly mixed acquisition DNA is mixed
Close liquid；
2. the pallet of above-mentioned molecular sieve bottom of the pillar is pushed up, make bottom lock, is placed in collecting pipe or centrifuge tube；So
The extract mixed liquor of DNA is added in above-mentioned DNA seperation films afterwards, is stopped 2-5 minutes, cotton-shaped genomic DNA and DNA separation
The combination of film specificity；
3. opening bottom tray, the biological macromolecule solns not adsorbed flow in collecting pipe or centrifuge tube, DNA seperation films
After cleaning solution, molecular sieve pillar is taken out, is inverted；
4. being fallen to DNA seperation films in new collecting pipe or centrifuge tube with external force, eluent is reused by DNA seperation films
On DNA dissolvings, collect genomic DNA up to sample.
For technique described above scheme, in the case of preferred, the disrupted sample cell is cracked by using DNA
Liquid, or the lysate containing protease k carry out.
For technique described above scheme, in the case of preferred, the main component of the elution solution be isopropanol or
Ethyl alcohol.
For technique described above scheme, in the case of preferred, the shaft is usually cylindrical shape, is received to be inserted in
It is used in collector or centrifuge tube；In the case of being more highly preferred to, the columnar high 5-12mm of shaft size, diameter 6-8mm；
Technique described above scheme is also set up in the case of preferred at the top of the shaft of above-mentioned molecular sieve pillar
There is top frame, in the case of being more highly preferred to, molecular sieve pillar shaft handle can also be increased on the frame of top；The top
The purpose of setting of frame is that molecular sieve pillar is fixed on collecting pipe or centrifuge tube, plays certain supporting function；Described point
Son sieves the setting purpose of pillar shaft handle in order to quickly and easily take out molecular sieve pillar.
Technique described above scheme is also set up in the case of preferred in the shaft bottom of above-mentioned molecular sieve pillar
There is molecular sieve pillar bottom tray；In the case of being more highly preferred to, handle can also be also set up in bottom tray；The molecular sieve
The purpose of setting of bottom of the pillar pallet is by shaft bottom lock, when DNA lysates being allowed to stop certain in molecular sieve pillar
Between, increase the binding time of DNA and DNA seperation films, to increase the extraction efficiency of DNA；The setting of the bottom tray handle
Purpose allows solution to flow out naturally to very easily open closed pallet.
The DNA seperation films have the hole flowed freely under gravity convenient for solution, the hole
Shape size does not limit, and in the case of preferred, the present invention at least enumerates following two kinds of situations, such as：DNA seperation films are that hole is
" v " type groove that the circular piece of 0.5-2.5 millimeters of apertures of diameter or the periphery of disk have number different, the seam of " v " type groove
Gap width is 0.5-2.5 millimeters；Those skilled in the art can select one or two kinds of hole in above two hole
Combination arrangement, such as：It is evenly arranged on disk on aperture or disk and is provided centrally with aperture, be around placed with 2-
There is no aperture among 20 " v " type grooves or disk, but periphery there are 2-20 " v " type grooves.
In the case of preferred, when the hole is 0.5-2.5 millimeters of apertures of diameter, aperture is to be evenly spaced in
In DNA seperation films, quantity is 5~100；
For technique described above scheme, in the case of preferred, the material of the DNA seperation films is the nitric acid of silanization
Aromatic polyamide fibre or glass fiber material of the polyesteramide fibre or silanization of fibrous material or silanization etc.；
For technique described above scheme, in the case of preferred, for DNA seperation films described above, material is removed
Glass fibre can be cut into disk after high-temp sterilizing, except direct use, for cellulose nitrate and other fibers
(polyesteramide fibre, aromatic polyamide fibre) material, before use should its DNA seperation film upper surface layer with silanization (silica gel)
Processing, the method for processing are Ethoxysilane solution to be dropped in the surface of DNA seperation films, or impregnated with Ethoxysilane solution
Fibrous material, it is spare after vacuum drying；Wherein：The silanizing solution：TEOS (Ethoxysilane), absolute ethyl alcohol and 30% hydrogen
Ammonium hydroxide solution, according to 0.2：13.8：6 volume ratio mixing；Wherein 30% Ammonia is to use preceding dropwise addition.
For technique described above scheme, in the case of preferred, for silanization (silica gel) processing side described above
Method is to handle silanizing solution and fibrous material room temperature at least 3 hours.
Advantageous effect：
Molecular sieve pillar of the present invention can extract the genomic DNA of high quality in a short period of time, reduce
Operating time reduces operating cost.It is compared with Qiagen genome DNA extracting reagent kits (product identification 51104), single sample
Extraction have following advantage：
1) taking for this method is 12 minutes, and the used time of Qiagen kits is 25 minutes；
2) instrument needed for this method is simple, it is thus only necessary to a micro tube oscillator and thermostat (65 DEG C), and
The method of Qiagen DNA extractions not only needs above mentioned micro tube oscillator and thermostat to also need to micro high speed centrifugation
Machine or vacuum pump；
3) the DNA mass of this method extraction is high, and yield is big, the extraction to people's saliva sample DNA, and average dna yield is
2-3 times of Qiagen methods, the extraction for blood sample DNA, the quality and yield and Qiagen kit methods of acquisition are suitable.
In short, molecular sieve pillar using the present invention, the method for extracting nucleic acid of progress is easy to operate, does not need complexity
Instrument and equipment, at low cost, DNA output is high, high-quality.Within 20 cents of the cost of single sample, single operation can be
12 or more samples are carried out at the same time；And the total time of single operation is only 10-30 minutes.
Description of the drawings
Fig. 1 molecular sieve pillars of the present invention, wherein：1. the shaft of molecular sieve pillar；2. molecular sieve pillar shaft handle
Hand；3.DNA seperation films；4.DNA detaches membrane support；5. molecular sieve pillar upper edge frame；6. molecular sieve pillar bottom tray；7. bottom
Portion's tray handle.
Other stand frame shapes of Fig. 2 .DNA seperation films；
The DNA solution and the mixing of 1 μ L 10 × DNA sample sample solutions that Fig. 3 .8 μ L are purified from people's saliva sample, are added 1%
Ago-Gel in, the purity of electrophoretic analysis sample.L, DNA standard items；1, molecular sieve pillar separation of the present invention is pure
The sample of change；The sample that 2, Qiagen DNA extraction kits isolate and purify.
The DNA solution and the mixing of 1 μ L 10 × DNA sample sample solutions that Fig. 4 .8 μ L are purified from human blood sample, are added 1%
Ago-Gel in, the purity of electrophoretic analysis sample.L, DNA standard items；1, molecular sieve pillar separation of the present invention is pure
Change sample；2, Qiagen kits samples.
Specific implementation mode
Following nonlimiting examples can make those skilled in the art be more fully understood the present invention, but not with
Any mode limits the present invention.
Embodiment 1
The preparation method of molecular sieve pillar：
If it is glass fiber material, it can be cut into disk after high-temp sterilizing, directly used.
If it is cellulose nitrate, polyesteramide fibre or aromatic polyamide fibre material need to carry out silanization treatment first
Afterwards, then be cut into disk use.The flow of the silanization treatment is as follows：
1) silanizing solution, 0.2mL TEOS (Ethoxysilane), in addition 13.8mL absolute ethyl alcohols, use preceding dropwise addition are prepared
6mL30% Ammonias；
2) fibrous material is impregnated in the solution, or solution is added dropwise on fibrous material, incubation at room temperature is handled 5 hours；
3) material is placed in 110 DEG C of baking ovens and is dried, room temperature preservation is spare under aseptic condition.
Embodiment 2
1. the molecular sieve column minor structure used in the embodiment of the present invention 3~4 is as follows：
Including shaft and a kind of DNA seperation films of rapid extraction genomic DNA, genome can be quickly and efficiently extracted
DNA.The material of DNA seperation films used in embodiment 3~4 is the cellulose nitrate material of silanization.
The shaft is cylindrical shape；Shaft size high 5-12mm, diameter 6-8mm；Its bottom is provided with for holding DNA points
The pallet of X fonts holder and closed bottom from film；Handle is also set up on the pallet；Wherein, the holder can also be adopted
It can also with the style of similar Fig. 2.
There is the DNA seperation films circular piece for being hole for 0.5-2.5 millimeters of apertures of diameter, aperture to be evenly spaced in
In DNA seperation films, quantity is 5~100；
The DNA seperation films free-standing is placed on holder, and the DNA seperation films and shaft, can be with there are gap
It is isolated and fallen off to shaft top orientation (one direction) under slight outer force effect.
It is additionally provided with top frame at the top of the shaft of above-mentioned molecular sieve pillar, frame increases shaft handle on top；
It is additionally provided with molecular sieve pillar bottom tray in the shaft bottom of above-mentioned molecular sieve pillar；It is arranged in bottom tray
Handle.
2. the application method of the molecular sieve pillar in above-described embodiment 3~4 is as follows：
When for extracting genome DNA, DNA lysates, or the lysate containing protease k, disrupted sample are first used
Then the isopropanol of 1/3-1/2 volumes is added in cell, be uniformly mixed the extract mixed liquor for obtaining DNA；
The pallet of molecular sieve bottom of the pillar is pushed up, bottom lock is made, is placed in collecting pipe or centrifuge tube；Then will
The extract mixed liquor of DNA adds in DNA seperation films, stops 2-5 minutes, cotton-shaped genomic DNA and DNA seperation film specificity
Combination, open bottom tray, the biological macromolecule solns not adsorbed flow in collecting pipe or centrifuge tube, and DNA seperation films are again
After cleaning solution, molecular sieve pillar is taken out, is inverted, is fallen to DNA seperation films in new collecting pipe or centrifuge tube with external force,
Eluent is reused by DNA dissolvings, the collection in DNA seperation films；It can be obtained the genomic DNA of high yield, high-quality.Its
In, the main component of the elution solution is isopropanol or ethyl alcohol.
The DNA sample lysate is the guanidine hydrochloride of 4.5mM, the Tris-HCl of 50mmol, pH 6.0,3.0%
Triton-X100；
Cleaning solution is 10mmol Tris-HCl, 80% ethyl alcohol；
DNA eluents are 10mmolTris-HCl, 1mmol EDTA, pH8.0；
Embodiment 3
Molecular sieve pillar of the present invention uses example 1 --- the extraction purification of people's saliva DNA
Utilize molecular sieve pillar of the present invention and Qiagen DNA extraction kits while extraction purification people's saliva
DNA, all processes are completed in 10 minutes.Required instrument is 65 DEG C of thermostats and microcentrifugal tube shaker.
1. operating process
1.1. sampling：Saliva retains at least 30 seconds in mouth, spits in a clean 50mL centrifuge tube, is turned with pipettor
It moves in 300 μ L salivas to the microcentrifugal tube of 1.5mL；
1.2.Qiagen the operating process of blood DNA extracts kit is carried out in strict accordance with the guide for use of kit.With
Under be molecular sieve pillar of the present invention DNA extraction process.
1.3. be added into the microcentrifugal tube equipped with saliva sample 400 μ L sample dissociation liquid (guanidine hydrochloride of 4.5mM,
The Triton-X100 of the Tris-HCl of 50mmol, pH 6.0,3.0%), quick oscillation 10 seconds；
1.4. 12.5 μ L protease k solution, quick oscillation 10 seconds is added；
1.5. 5 minutes are kept the temperature at 65 DEG C, quick oscillation 10 seconds；
1.6. 500 μ L isopropanols are added, turn upside down centrifuge tube 3 times, stands 30 seconds at room temperature；
1.7. solution all in sample processing tube is poured into molecular sieve pillar；
1.8. it is washed twice with cleaning solution (10mmol Tris-HCl, 80% ethyl alcohol), every time 300 μ L, i.e., directly by cleaning solution
It is added in duckpin, it is allowed to flow through naturally；
1.9. duckpin is upside down in a clean DNA collecting pipe, duckpin is gently squeezed with pipettor gun head
Bottom makes it fall to the bottom of collecting pipe, be added 100 μ L DNA eluents (10mmolTris-HCl, 1mmol EDTA,
PH8.0), quick oscillation 30 seconds.
2. experimental result：
1. the measurement (Nanodrop) of DNA concentration
It is found by the measurement of Nanodrop, the DNA output that molecular sieve pillar of the present invention isolates and purifies is classical
20 times or more of Qiagen methods, as purity (ratio of 260nm with 280nm hygroscopicity values).
Fig. 3 is the DNA solution and 1 10 × DNA of μ L of 8 μ L purifying
Sample sample solution mixes, and is added in 1% Ago-Gel, the purity of electrophoretic analysis sample.L, DNA standard items；
1, the sample that molecular sieve pillar of the present invention isolates and purifies；The sample that 2, Qiagen DNA extraction kits isolate and purify.
The DNA that molecular sieve pillar of the present invention isolates and purifies is through agarose gel electrophoresis (Fig. 3) display, DNA purity
Height does not see the impurity such as RNA.
Embodiment 4
Molecular sieve pillar of the present invention uses example 2 --- the extraction purification of human blood DNA
While Qiagen blood DNAs extracts kit (kit number 51104) is used, molecular sieve pillar of the present invention,
DNA is extracted from the whole blood that 100 μ L EDTA are handled.
1. operating process
1.1. sampling：It collects in the microcentrifugal tube that handle to EDTA of about 50 μ L whole bloods, takes 100 μ L to newly micro
In centrifuge tube；
1.2.Qiagen the operating process of blood DNA extracts kit and third party's kit makes in strict accordance with kit
It is carried out with guide.It is the DNA extraction process of molecular sieve pillar of the present invention below.
1.3. be added 300 μ L samples lysates (guanidine hydrochloride of 4.5mL, the Tris-HCl of 50mmol, pH 6.0,3.0%
), Triton-X100 quick oscillation 10 seconds；
1.4. 50 μ L protease k solution, quick oscillation 10 seconds is added；
1.5. 5 minutes are kept the temperature at 65 DEG C, quick oscillation 10 seconds；
1.6. 500 μ L isopropanols are added, turn upside down centrifuge tube 3 times, stands 30 seconds at room temperature；
1.7. the solution that sample collection is all from pours into molecular sieve pillar of the present invention；
1.8. it is washed twice with cleaning solution (10mmolTris-HCl, 80% ethyl alcohol), every time 300 μ L, i.e., directly by cleaning solution
It is added in duckpin, it is allowed to flow through naturally；
1.9. duckpin is upside down in a clean DNA collecting pipe, duckpin is gently squeezed with pipettor gun head
Bottom makes it fall to the bottom of collecting pipe, be added 100 μ L DNA eluents (10mmol Tris-HCl, 1mmol EDTA,
PH 8.0), quick oscillation 30 seconds.
2. experimental result：
The measurement (Nanodrop) of 2.1DNA concentration
It is found by the measurement of Nanodrop, the DNA output and classics that molecular sieve pillar of the present invention isolates and purifies
Qiagen methods obtain it is suitable, purity (260nm and 280nm water suction angle value ratio) also close to.
2.2 agarose gel electrophoresis (Fig. 4)
The blood sample of 10 different peoples of method pair of Fig. 4 present invention carries out DNA extractions, the use of sample size is 50 μ L,
After the completion of elution, DNA concentration and purity are measured with nanodrop.8 μ LDNA samples are taken to analyze sample using agarose gel electrophoresis
Purity.The DNA that molecular sieve pillar of the present invention isolates and purifies shows that DNA purity is higher, does not have through agarose gel electrophoresis
See the impurity such as RNA.
For any person skilled in the art, without departing from the scope of the technical proposal of the invention, all
Many possible changes and modifications are made to technical solution of the present invention using the technology contents of the disclosure above, or are revised as equivalent
The equivalent embodiment of variation.Therefore, every content without departing from technical solution of the present invention, according to the technical essence of the invention to
Any simple modifications, equivalents, and modifications that upper embodiment is done should all still fall within the range of technical solution of the present invention protection
It is interior.
Claims (10)
1. a kind of method of rapid extraction genomic DNA, it is characterised in that：The method includes using following molecular sieve pillars
Step：
The molecular sieve column attached bag includes shaft and DNA seperation films；The shaft bottom is provided with for holding DNA seperation films
The pallet of holder and closed bottom；The DNA seperation films have the hole flowed freely under gravity convenient for solution；
The DNA seperation films free-standing is placed on holder, and there are gaps for the DNA seperation films and shaft；
Method and step using above-mentioned molecular sieve pillar rapid extraction genomic DNA includes：
1. then the isopropanol of 1/3-1/2 volumes is added in disrupted sample cell, it is uniformly mixed the extract mixed liquor for obtaining DNA；
2. the pallet of above-mentioned molecular sieve bottom of the pillar is pushed up, make bottom lock, is placed in collecting pipe or centrifuge tube；Then will
The extract mixed liquor of DNA adds in above-mentioned DNA seperation films, stops 2-5 minutes, and cotton-shaped genomic DNA and DNA seperation films are special
Anisotropic combination；
3. opening bottom tray, the biological macromolecule solns not adsorbed flow in collecting pipe or centrifuge tube, and DNA seperation films pass through again
After over cleaning liquid, molecular sieve pillar is taken out, is inverted；
4. being fallen to DNA seperation films in new collecting pipe or centrifuge tube with external force, reusing eluent will be in DNA seperation film
DNA dissolvings, the genomic DNA for collecting sample to obtain the final product.
2. the method for rapid extraction genomic DNA according to claim 1, it is characterised in that：The disrupted sample is thin
Born of the same parents are by using DNA lysates, or the lysate progress containing protease k.
3. the method for rapid extraction genomic DNA according to claim 1, it is characterised in that：The master of the elution solution
It is isopropanol or ethyl alcohol to want ingredient.
4. the method for rapid extraction genomic DNA according to claim 1, it is characterised in that：In the molecular sieve pillar
Shaft at the top of be additionally provided with top frame；The shaft handle of molecular sieve pillar is additionally provided on the frame of top.
5. the method for rapid extraction genomic DNA according to claim 1, it is characterised in that：In above-mentioned molecular sieve pillar
Shaft bottom be additionally provided with molecular sieve pillar bottom tray；It is additionally provided with handle in the bottom tray.
6. the method for rapid extraction genomic DNA according to claim 1, it is characterised in that：The DNA seperation film bands
It is the aperture of 0.5-2.5 millimeters of diameter to have hole, or " v " type groove for being 0.5-2.5 millimeters with gap width.
7. the method for rapid extraction genomic DNA according to claim 6, it is characterised in that：The aperture is uniformly to arrange
For cloth in DNA seperation films, quantity is 5~100.
8. the method for rapid extraction genomic DNA according to claim 1, it is characterised in that：The material of the DNA seperation films
Matter is the cellulose nitrate material or the polyesteramide fibre of silanization or the aromatic polyamide fibre or glass of silanization of silanization
Fibrous material.
9. the method for rapid extraction genomic DNA according to claim 1, it is characterised in that：The DNA Separation membranes
Diameter is slightly less than internal diameter 1mm of shaft or so.
10. the method for rapid extraction genomic DNA according to claim 1, it is characterised in that：The shaft is cylinder
Shape, the columnar high 5-12mm of shaft size, diameter 6-8mm.
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| WO2011122841A2 (en) * | 2010-04-02 | 2011-10-06 | 주식회사 인트론바이오테크놀로지 | Column for extracting biochemical materials from biological sample |
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