CN110283814A - A kind of magnetic bead aaerosol solution and its application method - Google Patents
A kind of magnetic bead aaerosol solution and its application method Download PDFInfo
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- CN110283814A CN110283814A CN201910595352.9A CN201910595352A CN110283814A CN 110283814 A CN110283814 A CN 110283814A CN 201910595352 A CN201910595352 A CN 201910595352A CN 110283814 A CN110283814 A CN 110283814A
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- magnetic bead
- aaerosol solution
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- dna
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N15/1006—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
- 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
Abstract
The invention discloses a kind of magnetic bead aaerosol solution and its application methods, are to solve the very different problem of the quality of existing domestic magnetic bead Nucleic acid purification kits.This product includes following raw material: the polyethylene glycol that the dispersing agent and volume fraction that the biological buffer of final concentration of 0.1-2mol/L, the silicono magnetic bead of final concentration of 10-500ng/ μ L, the sodium chloride of final concentration of 1-20mol/L, volume fraction are 0.1-5% are 10-50%, and the pH value of magnetic bead aaerosol solution is 6-9.This product is cheap, the comparison highest XP magnetic bead of occupation rate of market, and under identical purifying ratio, the purification and recovery rate of this product is proximate to it；Under identical purifying ratio, the recycling clip size of this product is consistent with its；This product only uses magnetic frame, therefore compared to other conventional methods, flux is significantly improved；This product is easy to operate, not high to instrument requirements, and the operating time is short.
Description
Technical field
The present invention relates to nucleic acid purifications and segment to screen field, specifically a kind of magnetic bead aaerosol solution.
Background technique
High throughput sequencing technologies (NGS) cover library construction, the sequencing of upper machine, data analysis, medicine four modules of interpretation.Its
Chinese library building is experiment (WET LAB) end in the foundation stone and high throughput sequencing technologies of high throughput sequencing technologies.
Library construction is the following steps are included: nucleic acid fragment (chemical digestion or physics interrupt), end repair reaction, segment
Screening adds A to react with purifying, end, segment screening is reacted with purifying, jointing, segment screening is enriched with purifying, PCR amplification
Reaction, PCR product purifying, library quality inspection, (following processes be related to probe capture just need to carry out) hybrid capture, again PCR
The processes such as amplification enrichment reaction, PCR product purifying, library quality inspection.From the foregoing, it will be observed that being related to a large amount of core in library construction process
The work of acid purifying and segment screening.
Have gel extraction method for the conventional method that nucleic acid purification and segment screen and be centrifuged column method etc..Gel extraction method
Briefly steps are as follows: prepare Ago-Gel, point sample electrophoresis, gel extraction；The process is cumbersome, wants to electrophoretic buffer
Ask high, UV duration easily causes to damage to DNA.Centrifugal column method briefly steps are as follows: filter membrane absorption, washing lotion washing, dry filter membrane,
Elute nucleic acid；The process is cumbersome, and the rate of recovery is generally relatively low, cannot achieve high throughput.
The modernism screened for nucleic acid purification and segment has paramagnetic particle method.Most representative magnetic bead product surely belongs to Germany
The Agencourt AMPure XP of BeckMan company is a kind of magnetic bead nucleic acid purification examination that can obtain high quality DNA product
Agent box.Also there is the similar reagents box of Domestic Product on the market, but quality is very different, people are also carrying out grinding for related fields
Study carefully.
Summary of the invention
The embodiment of the present invention is designed to provide a kind of magnetic bead aaerosol solution, mentioned above in the background art to solve
Problem.
To achieve the above object, the embodiment of the present invention provides the following technical solutions:
A kind of magnetic bead aaerosol solution, including following raw material: the biological buffer of final concentration of 0.1-2mol/L, final concentration of
The silicono magnetic bead of 10-500ng/ μ L, the sodium chloride of final concentration of 1-20mol/L, volume fraction be 0.1-5% dispersing agent and
Volume fraction is the polyethylene glycol of 10-50%, and the pH value of magnetic bead aaerosol solution is 6-9.
As further embodiment of the embodiment of the present invention: dispersing agent uses polysorbas20, and biological buffer uses trihydroxy methyl
Aminomethane, market are easily bought, and using effect is good.
As further embodiment of the embodiment of the present invention: the relative molecular weight of polyethylene glycol is 1000-10000, can be incited somebody to action
Other solutes sufficiently dissolve, and using effect is good.
The magnetic bead aaerosol solution carries out the application method of DNA purifying, the specific steps are as follows:
Step 1 takes out magnetic bead aaerosol solution from 2-8 DEG C of refrigerator, is incubated for 25-35 minutes at room temperature；
Sample of nucleic acid to be purified is added in magnetic bead aaerosol solution, is then uniformly mixed, is incubated at room temperature by step 2
It 2-10 minutes, stands until liquid clarification, isolated supernatant and precipitating；
Precipitating is transferred in new test tube and is cleaned 2-3 times with ethyl alcohol by step 3, abandons raffinate, dries magnetic bead, do not do
It splits, then eluted dna, is stored at room temperature 2-6 minutes, is then placed on magnetic frame, the DNA of obtained supernatant i.e. after purification is produced
Object.
As further embodiment of the embodiment of the present invention: magnetic bead aaerosol solution is mixed using preceding using vortex vortex mixer
It is even, convenient for obtaining the high DNA product of purification degrees.
As further embodiment of the embodiment of the present invention: the volume fraction of ethyl alcohol is 80% in step 3.
The magnetic bead aaerosol solution carries out the application method of DNA fragmentation screening, the specific steps are as follows:
Step 1 takes out magnetic bead aaerosol solution from 2-8 DEG C of refrigerator, is incubated for 25-35 minutes at room temperature；
Sample of nucleic acid to be purified is added in magnetic bead aaerosol solution, is then uniformly mixed, is incubated at room temperature by step 2
It 2-10 minutes, stands until liquid clarification, isolated supernatant and precipitating；
Precipitating is transferred in new test tube and is added bead suspension, mixes and be incubated at room temperature 2- by step 3
It 10 minutes, stands up to clarification, supernatant is transferred in new test tube to and is added magnetic bead aaerosol solution, mix and in room temperature
Lower incubation 2-10 minutes, then cleaned 2-3 times with ethyl alcohol, raffinate is abandoned, dries magnetic bead, not dry and cracked, then eluted dna, is stored at room temperature
It 2-6 minutes, is then placed on magnetic frame, the DNA product of obtained supernatant i.e. after purification.
As further embodiment of the embodiment of the present invention: using low salt solutions or nuclease-free water eluted dna in step 3.
Compared with prior art, the beneficial effect of the embodiment of the present invention is:
This product uses the magnetic bead with silicono, and cheap, the highest XP magnetic bead of occupation rate of market, 1ml price exists
150-200 yuans, under this product batch production, price can realize a quarter of only XP magnetic bead price；
This product compares the highest XP magnetic bead of occupation rate of market, under identical purifying ratio, the purification and recovery rate of this product with
It is close；Under identical purifying ratio, the recycling clip size of this product is consistent with its；
This product only uses magnetic frame (16 holes or 96 holes), therefore compared to other conventional methods, flux has been obtained significantly
It improves；
This product is easy to operate, not high to instrument requirements, and the operating time is short, has a extensive future.
Detailed description of the invention
Fig. 1 is that magnetic bead aaerosol solution and XP product carry out comparative result figure of the wheel purifying in 0.5-1.2 times of additional amount.
Fig. 2 is that magnetic bead aaerosol solution and XP product carry out comparative result figure of the wheel purifying in 1.8 times of additional amounts.
Fig. 3 is first comparative result figure of the magnetic bead aaerosol solution from the progress two-wheeled purifying of XP product in different additional amounts.
Fig. 4 is second comparative result figure of the magnetic bead aaerosol solution from the progress two-wheeled purifying of XP product in different additional amounts.
Fig. 5 is third comparative result figure of the magnetic bead aaerosol solution from the progress two-wheeled purifying of XP product in different additional amounts.
Specific embodiment
The technical solution of the patent is explained in further detail With reference to embodiment.
Embodiment 1
A kind of magnetic bead aaerosol solution, including following raw material: the trishydroxymethylaminomethane of final concentration of 0.2mol/L, end are dense
Degree is the dispersing agent and volume that the silicono magnetic bead, the sodium chloride of final concentration of 10mol/L, volume fraction of 80ng/ μ L are 2.3%
The polyethylene glycol that score is 30%, the pH value of magnetic bead aaerosol solution are 6-9.
The magnetic bead aaerosol solution carries out the application method of DNA purifying, the specific steps are as follows:
1. magnetic bead solution is taken out from 5 DEG C of refrigerators, it is incubated for 30 minutes at room temperature.
2. please be vortexed mixing before use.
3. being added sample of nucleic acid to be purified into bead suspension, according to sample, the volume ratio of magnetic bead is the progress of table 1
Purifying；
Table 1
| Sample | 1 | 1 | 1 | 1 | 1 |
| Magnetic bead | 0.5× | 0.8× | 1× | 1.2× | 1.8× |
4. after being added, being vortexed and mixing, be incubated at room temperature 2-10 minutes.
5. being statically placed in magnetic frame to clarify up to liquid, supernatant is abandoned.
6. cleaning twice using the ethyl alcohol that volume fraction is 80%, raffinate is abandoned.
7. magnetic bead is dried, it is not dry and cracked, using nuclease-free water eluted dna, it is stored at room temperature 3 minutes.
8. placing on magnetic frame, supernatant is DNA product after purification.
The magnetic bead aaerosol solution carries out the application method of DNA fragmentation screening, the specific steps are as follows:
1. magnetic bead solution is taken out from 4 DEG C of refrigerators, it is incubated for 30 minutes at room temperature.Please be vortexed mixing before use.
2. sample of nucleic acid to be purified is added into bead suspension, according to the additional amount of 2 first round of table magnetic bead solution
It is operated with the additional amount of the second wheel magnetic bead solution；
Table 2
In table 2 × indicate nucleic acid samples volume.If sample volume is 50 μ L, for the first, then first round magnetic bead
Solution is 0.8 × 50 μ L=40 μ L using volume；Second wheel magnetic bead solution is 0.2 × 50 μ L=10 μ L using volume.
3. after the first round is added, being vortexed and mixing, be incubated at room temperature 8 minutes.
4. being statically placed in magnetic frame to clarify up to liquid, supernatant is transferred in new pipe, and the required magnetic bead of the second wheel is added
Volume is vortexed and mixes, is incubated at room temperature 6 minutes.
5. twice using ethyl alcohol cleaning, abandoning raffinate.
6. magnetic bead is dried, it is not dry and cracked.Using nuclease-free water eluted dna, it is stored at room temperature 4 minutes.
7. placing on magnetic frame, supernatant is DNA product after purification.
The product of embodiment 1 and XP magnetic bead are subjected to DNA purifying and segment is screened, takes pictures to result, as a result sees figure
1-5, from Fig. 1-5 as can be seen that this product and the effect of XP magnetic bead it is roughly the same.
The working principle of the embodiment of the present invention is: having the magnetic bead of silicono functional group by outer surface, in higher concentration
Polyethylene glycol and NaCl buffer in form bead suspension.DNA sample is added into bead suspension, and conformation changes therewith
Become, negatively charged phosphate group is largely exposed to outside, " electric bridge " is formed by Na+ and carboxyl, so that DNA is by Specific adsorption
To the magnetic bead surfaces with silicono, therefore, DNA sample by specificity and can select in the bead suspension under different proportion
Selecting property filters out required DNA fragmentation.Since magnetic bead also has superparamagnetism, the magnetic bead of DNA can will be combined in externally-applied magnetic field
It is aggregated, is discarded supernatant under effect, cleaned with ethyl alcohol and dried magnetic bead (not dry and cracked), then with low salt solutions or nuclease-free water
Eluted dna, eluted product is with high purity and is free of other impurity.It is enterprising that this method can be applied to manual or mating self-reacting device
Row operation.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.No
It should treat any reference in the claims as limiting the claims involved.
In addition, it should be understood that although this specification is described in terms of embodiments, but not each embodiment is only wrapped
Containing an independent technical solution, this description of the specification is merely for the sake of clarity, and those skilled in the art should
It considers the specification as a whole, the technical solutions in the various embodiments may also be suitably combined, forms those skilled in the art
The other embodiments being understood that.
Claims (8)
1. a kind of magnetic bead aaerosol solution, which is characterized in that including following raw material: the biological buffer of final concentration of 0.1-2mol/L
Agent, the silicono magnetic bead of final concentration of 10-500ng/ μ L, final concentration of 1-20mol/L sodium chloride, volume fraction 0.1-
5% dispersing agent and volume fraction is the polyethylene glycol of 10-50%, and the pH value of magnetic bead aaerosol solution is 6-9.
2. magnetic bead aaerosol solution according to claim 1, which is characterized in that the dispersing agent uses polysorbas20, and biology is slow
Electuary uses trishydroxymethylaminomethane.
3. magnetic bead aaerosol solution according to claim 1 or 2, which is characterized in that the relative molecular weight of the polyethylene glycol
For 1000-10000.
4. a kind of magnetic bead aaerosol solution a method according to any one of claims 1-3 carries out the application method of DNA purifying, feature exists
In, the specific steps are as follows:
Step 1 takes out magnetic bead aaerosol solution from 2-8 DEG C of refrigerator, is incubated for 25-35 minutes at room temperature；
Sample of nucleic acid to be purified is added in magnetic bead aaerosol solution, is then uniformly mixed, is incubated at room temperature 2-10 by step 2
Minute, it stands until liquid clarification, isolated supernatant and precipitating；
Precipitating is transferred in new test tube and is cleaned 2-3 times with ethyl alcohol by step 3, abandons raffinate, then eluted dna, room temperature are quiet
It sets 2-6 minutes, is then placed on magnetic frame, the DNA product of obtained supernatant i.e. after purification.
5. the application method that magnetic bead aaerosol solution according to claim 4 carries out DNA purifying, which is characterized in that the magnetic
Pearl aaerosol solution is uniformly mixed using preceding using vortex vortex mixer.
6. the application method that magnetic bead aaerosol solution according to claim 4 or 5 carries out DNA purifying, which is characterized in that described
The volume fraction of ethyl alcohol is 80% in step 3.
7. a kind of magnetic bead aaerosol solution a method according to any one of claims 1-3 carries out the application method of DNA fragmentation screening, feature
It is, the specific steps are as follows:
Step 1 takes out magnetic bead aaerosol solution from 2-8 DEG C of refrigerator, is incubated for 25-35 minutes at room temperature；
Sample of nucleic acid to be purified is added in magnetic bead aaerosol solution, is then uniformly mixed, is incubated at room temperature 2-10 by step 2
Minute, it stands until liquid clarification, isolated supernatant and precipitating；
Precipitating is transferred in new test tube and is added bead suspension by step 3, is mixed and is incubated at room temperature 2-10 points
Clock is stood up to clarification, and supernatant is transferred in new test tube to and is added magnetic bead aaerosol solution, is mixed and is incubated at room temperature
It educates 2-10 minutes, then is cleaned 2-3 times with ethyl alcohol, abandon raffinate, then eluted dna, is stored at room temperature 2-6 minutes, is then placed within magnetic
On power frame, the DNA product of obtained supernatant i.e. after purification.
8. the application method that magnetic bead aaerosol solution according to claim 7 carries out DNA fragmentation screening, which is characterized in that institute
It states in step 3 using low salt solutions or nuclease-free water eluted dna.
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| CN201910595352.9A CN110283814A (en) | 2019-07-03 | 2019-07-03 | A kind of magnetic bead aaerosol solution and its application method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111197043A (en) * | 2020-01-15 | 2020-05-26 | 深圳海普洛斯医学检验实验室 | Magnetic bead sorting solution for DNA separation and preparation method thereof |
| CN114213492A (en) * | 2021-09-23 | 2022-03-22 | 武汉奥科鼎盛生物科技有限公司 | DNA purification reagent, preparation method thereof and DNA purification method |
| CN114480369A (en) * | 2021-12-24 | 2022-05-13 | 广东润鹏生物技术有限公司 | Magnetic bead preservation solution, preparation method and magnetic bead product |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109735532A (en) * | 2018-12-30 | 2019-05-10 | 北京优迅医学检验实验室有限公司 | A kind of bead suspension and its application for nucleic acid purification and screening |
-
2019
- 2019-07-03 CN CN201910595352.9A patent/CN110283814A/en not_active Withdrawn
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109735532A (en) * | 2018-12-30 | 2019-05-10 | 北京优迅医学检验实验室有限公司 | A kind of bead suspension and its application for nucleic acid purification and screening |
Non-Patent Citations (1)
| Title |
|---|
| MICHAEL BAYM等: "Inexpensive Multiplexed Library Preparation for Megabase-Sized Genomes", 《PLOS ONE》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111197043A (en) * | 2020-01-15 | 2020-05-26 | 深圳海普洛斯医学检验实验室 | Magnetic bead sorting solution for DNA separation and preparation method thereof |
| CN114213492A (en) * | 2021-09-23 | 2022-03-22 | 武汉奥科鼎盛生物科技有限公司 | DNA purification reagent, preparation method thereof and DNA purification method |
| CN114480369A (en) * | 2021-12-24 | 2022-05-13 | 广东润鹏生物技术有限公司 | Magnetic bead preservation solution, preparation method and magnetic bead product |
| CN114480369B (en) * | 2021-12-24 | 2022-12-27 | 广东润鹏生物技术有限公司 | Magnetic bead preservation solution, preparation method and magnetic bead product |
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Application publication date: 20190927 |