CN110106169A - A method of improving trace dna capture rate in cell-free body fluid - Google Patents
A method of improving trace dna capture rate in cell-free body fluid Download PDFInfo
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Abstract
The present invention provides a kind of method for improving trace dna capture rate in cell-free body fluid, by protein cleavage, crosses column and combines, cleaning and elution and etc. realization.Conventional protein cleavage formula and conventional nucleic acid can be used to cross column combination purification process for the method for the present invention, crucial centrifugation need to only be added and go liquid step, without using large volume combination liquid, without using magnetic bead, it is easy to operate, additional skill is not needed, reagent cost is low, compared with general histocyte nucleic acid extraction kit, trapping nucleic acids rate can be improved to 3-8 times.It is similar to dedicated blood plasma free serum nucleic acid extraction kit obtained by nucleic acid extraction, it is a kind of economic and effective method.
Description
Technical field
The invention belongs to field of biotechnology, are related to a kind of method for improving trace dna capture rate in cell-free body fluid,
For improving trapping nucleic acids rate when enrichment and purification of nucleic acid from cell-free body fluid, especially suitable for rich from serum or blood plasma
The occasion of collection and purifying minim DNA.
Background technique
Liquid biopsy is the hot spot of current biological medical domain research.Liquid biopsy is primarily referred to as the serum from living body, blood
Trace dna substance is extracted in slurry, cerebrospinal fluid etc. are cell-free body fluid, further detection is for prompting disease.Trace dna substance master
Refer to DNA, but also includes all kinds of RNA.It is mainly that dead cell is released from cell-free serum and the extracted nucleic acid substances of blood plasma
It puts, is recycled in hematological system in vivo, can be described as free nucleic acid, main ingredient is referred to as dissociative DNA.Dissociative DNA, which has, to wither
The feature for dying cell institute released dna, in about 170bp and its position of integral multiple length when showing as the DNA electrophoresis of institute's extraction purification
It is equipped with product band, and the large fragment of less 1000bp or more.Therefore, DNA is extracted compared to thin in cell-free serum or blood plasma
The genomic DNA that born of the same parents extract has significant difference.
When body is there are disease, the releasable nucleic acid substances of cell death caused by disease, free nucleic acid carries disease at this time
The information of disease.The meaning of liquid biopsy is exactly based on to being extraction purification in main syllabus target body fluid with cell-free serum and blood plasma
Nucleic acid substances detected, to achieve the purpose that auxiliary diagnosis disease, and be possible in the future in certain diseases as examining
Disconnected standard.
Nucleic acid substances in cell-free serum or blood plasma are deniers.Serum or blood plasma are as nucleic acid extraction purifying pair
As the volume handled needed for initial is likely present shadow not yet explicitly far beyond extraction tissue Shi Wei great in serum plasma
Ring the composition of nucleic acid absorption capture, this strong influence extraction efficiency of nucleic acid.It is generally used for histocyte nucleic acid extraction
Kit can not almost obtain effective extraction.There are a small number of dedicated commercial kits to can provide more efficient extraction at present,
In with the QIAamp Circulating Nucleic Acid reagents series box effect of German QIAGEN company be it is excellent.QIAGEN
Company's kit was mainly column purification type, which crosses column combination liquid using large volume and remove dilution large volume serum or blood plasma
Influence to extraction efficiency can be successfully acquired trace dna, but (50 extractions of price at present need nearly 10,000 yuan of people to higher cost
People's coin).Another magnetic bead absorption method has more company to provide product, but paramagnetic particle method in large volume liquid to the suction of trace dna
Attached capture rate is unsatisfactory, especially when free nucleic acid amount is less.Compared with crossing column method, paramagnetic particle method trapping nucleic acids rate is inclined
It is low, nucleic acid substances are inevitably lost when cleaning magnetic bead.In addition, paramagnetic particle method to magnetic bead particles diameter, suspended dispersed degree,
Homogeneity etc. requires height, and kit cost is still higher.Paramagnetic particle method, which is extracted, requires also higher, extraction to the operation technique of operator
Effect is also not sufficiently stable.
Summary of the invention
The object of the present invention is to provide a kind of methods for improving trace dna capture rate in cell-free body fluid, including albumen to split
Solution, cross column combine, cleaning and elution and etc., wherein key be protein cleavage and cross column combine between following steps are added:
(1) low dielectric constant solvent of 0.5-3 times of volume has been added in the centrifuge tube of crack protein body fluid by Xiang Shengyou, mixes；
(2) high speed is centrifuged at normal temperature；
(3) with direct pouring liquid, perhaps absorb liquid or be centrifuged again after toppling over the method removal liquid absorbed again at
Part；
(4) column combination liquid is crossed with nucleic acid redissolve adherent nucleic acid.
It is realized especially by following steps:
(1) cell-free body fluid is added in the plastic centrifuge tube of high-molecular compound material, adds common Proteinase K and egg
White denaturant protein lysate as the main component, is incubated under Proteinase K appropriate working temperature, makes protein macromolecule in liquid
Cracking, purpose make do not occur a large amount of albumen precipitations when subsequent high speed centrifugation, and the trace dna in liquid removes core in Proteinase K
It is completely saved under the protection of sour enzyme function.
Cell-free body fluid, the body fluid comprising human body and animal, comprising blood plasma, serum, amniotic fluid, cerebrospinal fluid, hydrothorax, ascites,
Hydropericardium, sinus cavities hydrops, synovial bursa hydrops, lactiferous ducts liquid and aqueous humor.
Used centrifuge tube is high-molecular compound material, comprising with polypropylene, polyethylene, polyvinyl chloride, polyphenyl second
Alkene, the plastic centrifuge tube that polycarbonate is main material.Preferably polypropylene material.Comprising sharp bottom, round bottom, square bottom cylinder
Plastic centrifuge tube.
Protein cleavage formula of liquid of the present invention must include two kinds of key components of Proteinase K and protein denaturant, this is
Known common prescription, solution pH value is best between 7.5-8, but pH value can also play protein cleavage effect between 7-9.
Protein lysate of the present invention also may include Tris, EDTA etc. other with chelation of metal ion, pH value buffer function
Conventional ingredients, be mainly used for adjusting pH value and chelated metal ions, be conventional method.
Proteinase K dosage is different with its protein content according to the serum or plasma volume of required processing in the protein lysate
Can be different, Proteinase K additional amount can be excessive, but cannot be insufficient.It is formed in serum (blood plasma) and protein lysate mixed
It closes in liquid, when Proteinase K content is 2ug/ microlitres, the blood plasma or haemocyanin cracking that can meet general protein content are required, right
It is enough in most blood plasma or serum.
Protein denaturant include mass percent be 0.2-20% lauryl sodium sulfate, the acetic acid of 0.5-8mol/L,
The guanidine hydrochloride of 0.9-8mmol/L, the guanidine thiocyanate of 0.9-8mmol/L, the urea of 4-8mmol/L and above-mentioned protein denaturant
The mixed liquor of precipitating is not precipitated.The above protein denaturant is known common prescription.
The temperature and time that protein cleavage of the present invention is incubated for according to handled sample size number can be different.Albumen
Enzyme K has certain activity between 30 DEG C and 80 DEG C, but relatively high with activity between 35 DEG C -60 DEG C, preferably with 55 DEG C
Incubation is preferred.When handled sample size less (such as < 100 microlitres), 35 DEG C or 80 DEG C incubations can reach crack protein
Effect, and incubation time 10 minutes.When handle sample size it is larger when (as > 100 microlitres) when, preferably with 50-60 DEG C it
Between be incubated for, incubation time should be greater than 1 hour, preferably 3 hours, also can be incubated overnight, but be not to be exceeded 16 hours.
(2) it crosses column to combine: after the completion of protein cleavage, original liquid volume 0.5 being added into centrifuge tube again to 3 times of volumes
Low dielectric constant solvent, in the state that centrifuge tube and centrifugal shaft form inclination angle, in the environment not less than 4 DEG C and no more than 35 DEG C
At a temperature of, middling speed or high speed centrifugation make in liquid trace dna applied solid in plastic centrifuge tube wall.It directly uncaps after centrifugation
Liquid is removed, be centrifuged again there are raffinate person and is removed liquid component in centrifuge tube with the method that pipettor absorbs liquid, mesh
It is to remove liquid feed in centrifuge tube.
Low dielectric constant solvent refers to that dielectric constant includes ethyl alcohol, isopropanol, second less than the solvent of pure water at 20 DEG C
Acid, acetone, n-hexyl alcohol and carbon tetrachloride.Preferably ethyl alcohol or isopropanol.It is of the present invention after the completion of protein cleavage, to splitting
Solve the solvent that low-k is added in liquid.The low dielectric constant solvent for addition should be sterling, i.e., content answer >
99%.Institute's additive amount according to the dielectric constant of solvent and the relative size difference of the dielectric constant of water and it is different, water is at 20 DEG C
Dielectric constant is 78, ethyl alcohol 24, and mass content of the ethyl alcohol in finally centrifugation solution should be greater than 60% at this time, and additional amount is general
It is 2 times of original solution volume.Isopropanol dielectric constant at 20 DEG C is 18, mass content of the isopropanol in finally centrifugation solution
It should be greater than 33%, quantity of isopropanol is generally 1 times of original solution volume.Acetone, n-hexyl alcohol, acetic acid and carbon tetrachloride are at 20 DEG C
When dielectric constant be respectively 21,13,6 and 2, additional amount is to be typically no less than 1.5 times, 0.75 times, 0.5 times of original solution volume
It can play a role with 0.5 times.
It is centrifuged at normal temperature after low dielectric constant solvent is added in cracking system liquid of the present invention, when comprising centrifugation
The environment temperature of centrifuge tube is room temperature, and temperature is between 4 DEG C and 35 DEG C, preferably 4 DEG C or 20 DEG C.Middling speed or high speed from
The heart.Middling speed or high speed centrifugation refer to centrifugal force in 1000 grams -30000 grams, play the role of making the adherent adherency of nucleic acid.It is preferred that being centrifuged
Power is 8000 grams, centrifugal force 40 minutes.
Centrifugal force is smaller, and the time needed for making nucleic acid applied solid is longer, when for the 30 degree of inclinations of 2ml centrifuge tube, when
Being centrifuged when 1000 grams of centrifugal force 2 hours can be securely adherent, 50 minutes when 4000 grams of centrifugal forces can be securely adherent, when
Need to only be centrifuged when 30000 grams of centrifugal force 10 minutes can be securely adherent.The position of centrifuge tube should make centrifuge tube major diameter side when centrifugation
Centrifugation inclination angle is formed to centrifugation axis direction, centrifugation inclination angle can be between 20-70 degree, preferably 30-40 degree horn shape state, purpose
For the distance for reducing traveling needed for nucleic acid in solution is attached at centrifugation tube wall, nucleic acid is made faster to be attached at centrifugation tube wall.Meanwhile core
Acid is attached at the side of centrifugation tube wall, is also beneficial to subsequent topple over or absorb liquid in the other side.
It by directly toppling over, or absorbs, or the method toppled over plus absorbed will be centrifuged liquid in pipe removal, residual liquid total amount
No more than 50 microlitres.
(3) routine is added into centrifuge tube and crosses column combination liquid, vortex oscillation mixes well.When vortex oscillation, in centrifuge tube
Liquid quickly rotates, and adherent liquid level increases, and the method that can be used centrifuge tube upright and be inverted intersection vortex oscillation makes to rotate liquid
Body touches all positions that centrifugation tube wall is possible to absorption nucleic acid.Vortex oscillation makes adherent nucleic acid be redissolved in column combination
In liquid.It is subsequent to cross nucleic acid purification post according to a conventional method, liquid amplifying nucleic acid is adsorbed in the silication adsorbed film of purification column, cleaning is added
Liquid centrifugation after, then with routine eluent by Nucleic Acid Elution, can be obtained the cell-free acid substance of center containing protein liquid of purifying.
Cross the combination liquid that column combination liquid is the low pH value with high salt applied to conventional pillar nucleic acid purification silication adsorbed film.It is known
Common formula includes sodium chloride, the lithium chloride, sodium molybdate, guanidine hydrochloride, sulphur (different sulphur) cyanic acid that molar concentration is between 2-8 moles
Guanidine, sodium salicylate, ammonium sulfate, sodium acetate, PEG 8000 and above compound of the mass percent between 5-15%
Mixed liquor.It can also be used what is be not publicly formulated in nucleic acid extraction kit in commodity market to cross column combination liquid.
The principle that the method for the invention can generate corresponding effect may is that the body fluid containing albumen such as blood plasma, serum are cracking
After albumen, in the presence of the low dielectric constant solvent of higher concentration, at normal temperature not less than 1000 grams of centrifugal force long periods from
The heart can make the nucleic acid being dispersed in liquid attaching be gathered in plastic centrifuge tube wall.And trace dna and plastic centrifuge tube wall are tight
When contiguity touching, it can be securely adsorbed in centrifugation tube wall, without occurring loosening and falling as a large amount of nucleic acid are adherent, this is not yet known
Discovery.Another possible principle is body fluid containing albumen after crack protein, there are still non-proteinaceous such as some lipids, these
Substance may be attached at plastic tube wall prior to trace dna, and forming a tunic can be by micro core when trace dna is fallen within thereon
Sour securely absorption.It is securely adsorbed, liquid portion can be removed as far as possible, this, which is just eliminated in former serum plasma, influences core by nucleic acid
Acid crosses the factor of column absorption, and the column combination liquid of crossing for redissolving trace dna is enable normally to play suction-operated.
The beneficial effects of the present invention are this method, and conventional protein cleavage formula and conventional nucleic acid can be used to cross column knot
Purification process is closed, crucial centrifugation need to only be added and go liquid step, without using large volume combination liquid, without using magnetic bead, operation
Simply, additional skill is not needed, reagent cost is low, compared with general histocyte nucleic acid extraction kit, can catch nucleic acid
The rate of obtaining is improved to 3-8 times.It is similar to dedicated blood plasma free serum nucleic acid extraction kit obtained by nucleic acid extraction, be a kind of economy again
Effective method.
Liquid biopsy such as in the future can large-scale application, trace dna extracting method also needs more economical in blood plasma or serum
And effective method.Our invention then provides a kind of very economic method, can be by shirtsleeve operation by common nucleic acid
Trapping nucleic acids rate of the reagent for when extracting trace dna from the body fluid such as blood plasma, serum is extracted to significantly improve.
For existing from the body fluid such as cell-free serum, blood plasma, cerebrospinal fluid when extraction purification trace dna, trapping nucleic acids rate
Not high perhaps higher cost or the unstable deficiency of effect, crack using by albumen in cell-free body fluid through conventional method
Afterwards, low dielectric constant solvent is added, is centrifuged at normal temperature, makes the adherent secure adhesion of nucleic acid in plastic centrifuge tube wall, be centrifuged
Column is crossed with nucleic acid and is combined by direct pouring liquid or the method absorbed, as far as possible removal liquid in pipe part immediately after
Liquid redissolves trace dna adherent in pipe, then the technical solution of column extraction is crossed with conventional method.The invention can use conventional reagent
Trapping nucleic acids rate when extracting trace dna in cell-free body fluid significantly improves, solve extracted from cell-free body fluid it is micro
Nucleic acid needs the problem of economic, effective, stable method, is conducive to the popularization and application of liquid biopsy.
Detailed description of the invention
Fig. 1 is 50ml polypropylene plastics material centrifuge tube used when purifying 6ml blood plasma.
Fig. 2 is to form fixed centrifugation inclination angle with centrifugation arbor when centrifuge tube is placed in centrifuge.
Fig. 3 is centrifuge tube frenulum pouring liquid upward.
Fig. 4 is that the vortex oscillation after crossing column combination liquid for redissolving adherent nucleic acid is added.
Specific embodiment
A specific embodiment of the invention is given below and is illustrated in conjunction with attached drawing.
Embodiment 1: the trapping nucleic acids rate that dissociative DNA is purified from larger amount of 6 milliliters of blood plasma is improved
As shown in fig. 1,6 milliliters of blood plasma are contained in the centrifuge tube of 50 milliliters of polypropylene materials of a round bottom in this example, should
Centrifuge tube has frenulum 1 to be connected with press cover 2, is provided with 6 milliliters of blood plasma 3.Proteinase K is added into centrifuge tube and protein denaturant is total
2 milliliters, add up to 8 milliliters.It is incubated for 3 hours in 55 degree of water-baths, cracks albumen sufficiently.After the completion of cracking, one times is added
Volume (8 milliliters) isopropanol forms 16 milliliters of liquid.It is put into the centrifugal hole of centrifuge such as Fig. 2, at this time 50 milliliters of centrifugations
The centrifugation inclination angle 6 that the major diameter 4 of pipe and the centrifugal shaft 5 of centrifuge are formed is 30 degree.In the centrifuge under 4 degrees Celsius, 1000 grams
Centrifugal force 2 hours.After the completion of centrifugation, centrifuge tube is taken out immediately, opens lid.It, will be from if Fig. 3 topples over and overturn centrifuge tube
Whole liquid 8 are all removed liquid in pipe by the opening 7 of centrifuge tube frenulum opposite side in heart pipe.It will fall to remove liquid again
Centrifuge tube be placed in former centrifuge, pays attention to being consistent when centrifuge tube frenulum direction must be with previous centrifugation.At 4 degrees Celsius, 1000
It is centrifuged 5 minutes under gram centrifugal force.After the completion of second is centrifuged, centrifuge tube is taken out immediately, it will with 200 microlitres of tip suction pipette heads
Residual liquid is absorbed in managing.When paying attention to absorbing liquid, centrifuge tube frenulum is kept upward, absorbing liquid at this time will not influence adherent core
Acid.After absorbing liquid, conventional nucleic acid is added into centrifuge tube and crosses totally 1500 microlitres of column combination liquid 9, is vibrated on turbula shaker
It mixes.When vortex oscillation, centrifugation liquid in pipe quickly rotates and is formed vortex, if Fig. 4 peripheral liquid 10 is on centrifugation tube wall
It rises, the method that can be used centrifuge tube upright and be inverted intersection vortex oscillation makes liquid level rotate to the institute that there may be adherent nucleic acid
There is centrifugation tube wall.Vortex oscillation mixes after five minutes, routinely cross column combine, cleaning and elution, can be obtained and extract in blood plasma
Free nucleic acid substance.Compared with the general method for extracting nucleic acid for not increasing step of the present invention, increase the method for the invention
Afterwards, the amount for obtaining free nucleic acid can increase to 3-8 times.
Embodiment 2: the trapping nucleic acids rate that dissociative DNA is purified from small amount of 450 microlitres of serum is improved
The container that this example contains serum is the cone bottom centrifuge tube of common 2 milliliters of polypropylene materials, and operational illustration yet can refer to reality
Apply the marked attached drawing of example 1.After 450 microlitres of serum are added in centrifuge tube, common Proteinase K and protein denaturant totally 150 is added
Microlitre, it is incubated for 3 hours in 55 degree of constant-temperature metal baths, cracks albumen in serum sufficiently, it is totally 600 micro- to be centrifuged liquid in pipe at this time
It rises.After the completion of cracking, 1200 microlitres of dehydrated alcohol is added, turns upside down 15 times and mixes well, it is totally 1800 micro- in pipe at this time
Rise liquid.Centrifuge tube is placed in the centrifuge that centrifugation inclination angle is 40 degree, at room temperature, centrifugal force is centrifugation 40 under 8000 grams
Minute.When paying attention to centrifugation, centrifuge tube pipe lid frenulum must partially upward, the label being positioned against in this, as centrifuge tube.It has been centrifuged
Cheng Hou takes out centrifuge tube immediately, opens lid, slowly overturns centrifuge tube, liquid is directly poured out from the opposite side of centrifuge tube frenulum,
And nozzle liquid is absorbed with dust-free paper, remove liquid.It adds 500 microlitres of common nucleic acid and crosses column combination liquid, vortex oscillation 5 is divided
Clock dissolves adherent nucleic acid sufficiently.It is conventional to cross column absorption, cleaning and elution, it can be obtained dissociative DNA.With do not increase institute of the present invention
The conventional method for stating raising trapping nucleic acids rate method is compared, and after increasing step of the present invention, dissociative DNA amount obtained increases
To 3-8 times.
Claims (10)
1. a kind of method for improving trace dna capture rate in cell-free body fluid, including protein cleavage are crossed column and are combined, clean and wash
Take off and etc., which is characterized in that following steps are added between protein cleavage and excessively column combine:
(1) low dielectric constant solvent of 0.5-3 times of volume has been added in the centrifuge tube of crack protein body fluid by Xiang Shengyou, mixes；
(2) high speed is centrifuged at normal temperature；
(3) with direct pouring liquid, perhaps absorb liquid or be centrifuged again after toppling over the method removal liquid absorbed again at
Part；
(4) column combination liquid is crossed with nucleic acid redissolve adherent nucleic acid.
2. a kind of method for improving trace dna capture rate in cell-free body fluid according to claim 1, which is characterized in that
It is realized by following steps:
(1) protein cleavage: cell-free body fluid is added in centrifuge tube, adds protein lysate incubation；
(2) after the completion of protein cleavage, it is molten to the low-k of 3 times of volumes again that original liquid volume 0.5 is added into centrifuge tube
Agent, in the state that centrifuge tube and centrifugal shaft form inclination angle, under the environment temperature not less than 4 DEG C and no more than 35 DEG C, middle height
Speed centrifugation makes trace dna applied solid in liquid directly uncap after centrifugation in plastic centrifuge tube wall and remove or absorb liquid,
It is centrifuged there are raffinate person and is removed liquid component in centrifuge tube again with the method that pipettor absorbs liquid, purpose is to make to be centrifuged
Liquid feed removes in pipe；
(3) be added into centrifuge tube it is conventional cross column combination liquid, vortex oscillation mixes well, when vortex oscillation, centrifugation liquid in pipe
Quickly rotation, adherent liquid level increase, keep centrifuge tube upright and be inverted intersect vortex oscillation method make rotating liquid touch from
Heart tube wall is possible to all positions of absorption nucleic acid, and vortex oscillation was redissolved in adherent nucleic acid in column combination liquid, after
It is continuous to cross nucleic acid purification post according to a conventional method, liquid amplifying nucleic acid is adsorbed in the silication adsorbed film of purification column, cleaning solution centrifugation is added
Afterwards, then with conventional eluent by Nucleic Acid Elution, it can be obtained the cell-free acid substance of center containing protein liquid of purifying.
3. a kind of method for improving trace dna capture rate in cell-free body fluid according to claim 2, which is characterized in that
The pH value of step (1) described protein lysate is between 7.5-8.
4. a kind of method for improving trace dna capture rate in cell-free body fluid according to claim 2, which is characterized in that
The temperature that step (1) protein cleavage is incubated for is but relatively high with activity between 35 DEG C -60 DEG C between 30 DEG C and 80 DEG C,
Preferably it is preferred with 55 DEG C of incubations.When handled sample size less (such as < 100 microlitres), 35 DEG C or 80 DEG C incubations are reachable
To the effect of crack protein, and incubation time 10 minutes.When processing sample size is larger (such as > 100 microlitres), preferably
To be incubated between 50-60 DEG C, incubation time should be greater than 1 hour, preferably 3 hours, also can be incubated overnight, but be not to be exceeded 16
Hour.
5. a kind of method for improving trace dna capture rate in cell-free body fluid according to claim 2, which is characterized in that
Step (2) low dielectric constant solvent refers to that at 20 DEG C dielectric constant is less than the solvent of pure water, comprising ethyl alcohol, isopropanol, acetic acid,
Acetone, n-hexyl alcohol and carbon tetrachloride；The low dielectric constant solvent for addition should be sterling, i.e. content answers > 99%.
6. a kind of method for improving trace dna capture rate in cell-free body fluid according to claim 2, which is characterized in that
It is centrifuged at normal temperature after low dielectric constant solvent is added in cracking system liquid described in step (2), centrifuge tube when comprising centrifugation
Environment temperature be room temperature, temperature is between 4 DEG C or 20 DEG C；The high speed centrifugation time, centrifugal force was 1000 between 10-120 minutes
Between grams -30000 grams.
7. a kind of method for improving trace dna capture rate in cell-free body fluid according to claim 2, which is characterized in that
Step (2), which passes through, directly topples over, or absorbs, or the method toppled over plus absorbed will centrifugation liquid in pipe removal.
8. a kind of method for improving trace dna capture rate in cell-free body fluid according to claim 2, which is characterized in that
It is the combination liquid with the low pH value with high salt of conventional pillar nucleic acid purification silication adsorbed film that step (3), which crosses column combination liquid,.
9. a kind of method for improving trace dna capture rate in cell-free body fluid according to claim 1 or 2, feature exist
In centrifuge tube major diameter direction and centrifugal force direction should form centrifugation inclination angle, and angle is between 20-70 degree；Centrifuge tube include sharp bottom,
Round bottom, square bottom cylindrical plastic centrifuge tube, centrifuge tube material main ingredient includes polypropylene, polyethylene, polyvinyl chloride, polyphenyl
Ethylene, polycarbonate.
10. a kind of method for improving trace dna capture rate in cell-free body fluid according to claim 6, feature exist
In centrifugal force is 8000 grams, centrifugal force 40 minutes.
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| CN201910451855.9A CN110106169A (en) | 2019-05-28 | 2019-05-28 | A method of improving trace dna capture rate in cell-free body fluid |
| PCT/CN2019/094963 WO2020237781A1 (en) | 2019-05-28 | 2019-07-07 | Method for increasing capture rate of trace nucleic acid in cell-free body fluid |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113046417A (en) * | 2021-03-25 | 2021-06-29 | 武汉吉诺百客医学科技有限公司 | Kit for extracting free DNA of blood plasma by paramagnetic particle method and use method thereof |
| CN113403395A (en) * | 2021-06-03 | 2021-09-17 | 南京世和基因生物技术股份有限公司 | Method and kit for extracting cfDNA of aqueous humor and application of kit in PVRL clinical auxiliary examination |
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Also Published As
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| WO2020237781A1 (en) | 2020-12-03 |
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