CN108517290A - A kind of five chamber nucleic acid-extracting apparatus and method based on immiscible phase interfacial tension - Google Patents
A kind of five chamber nucleic acid-extracting apparatus and method based on immiscible phase interfacial tension Download PDFInfo
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- CN108517290A CN108517290A CN201810548997.2A CN201810548997A CN108517290A CN 108517290 A CN108517290 A CN 108517290A CN 201810548997 A CN201810548997 A CN 201810548997A CN 108517290 A CN108517290 A CN 108517290A
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—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
A kind of five chamber nucleic acid-extracting apparatus and method based on immiscible phase interfacial tension, device includes sample cavity, sample cavity outlet passes sequentially through trapezoidal microchannel and is connected with the first immiscible phase chamber, washing cavities, the second immiscible phase chamber, elution chamber, and magnet is equipped with outside sample cavity, immiscible phase chamber, washing cavities or elution chamber；Method is that biological sample, cell pyrolysis liquid and nanometer magnetic bead are first injected into sample cavity, elution chamber is added in elution buffer, then washing cavities are added in cleaning solution, finally two immiscible phase chambers is given to fill immiscible phase；Magnet is rotated in sample chamber bottom outside, so that nanometer magnetic bead reacts to form magnetic bead nucleic acid complexes with biological sample, by magnet successively to the first immiscible phase chamber, washing cavities, the second immiscible phase chamber, elution chamber movement so that magnetic bead nucleic acid complexes enter elution chamber elution, and nucleic acid is discharged；The present invention realizes the rapid and convenientization extraction of nucleic acid, reduces the consumption of reagent sample, and reduce the loss of extraction process amplifying nucleic acid.
Description
Technical field
The present invention relates to the extraction purification technical fields of nucleic acid, and in particular to it is a kind of based on immiscible phase interfacial tension five
Chamber nucleic acid-extracting apparatus and method.
Background technology
Molecule diagnosis is mainly used in clinical diagnosis, such as tumour, infection, heredity using nucleic acid as detection object
Etc. various aspects, and " first step " of the nucleic acid extraction as molecular diagnostic assays, extraction rate and purity are most basic requirements.It is existing
The extracting method of modern nucleic acid mainly has phenol chloroform extraction method, centrifugal column extraction method, nanometer magnetic bead extraction method.Phenol chloroform extraction
The chemical precipitation methods such as method are most traditional nucleic acid purification methods, have a higher nucleic acid extraction rate, but operating process need it is anti-
Multiple precipitation, centrifugation, it is difficult to realize automation, and need to use toxic reagent, therefore gradually be eliminated.Then develop with from
Stem method is the solid phase carrier absorption method of representative, but its operating process is also required to high speed centrifugation repeatedly, will produce larger machinery
Power destroys the integrality of nucleic acid；It is not easy to high-throughput, automation mechanized operation simultaneously.Paramagnetic particle method nucleic acid extraction realizes micro ＆ nano technology
With merging for biotechnology, it is silicon dioxide coated by being carried out to magnetic bead, when nucleic acid is discharged from cell, nanometer magnetic bead and core
Acid specific binding, is formed magnetic bead-nucleic acid complexes, is done under the action of externally-applied magnetic field using the superparamagnetism of nanometer magnetic bead
Displacement can be such that compound is detached with other substances.However nanometer magnetic bead extraction method traditional at present, extraction step are：
Cell cracking-nanometer magnetic bead is combined-washing-elution with nucleic acid.Nucleic acid enriching purification time length (about 1h)；Secondly, it operated
Journey needs that the operations such as demagnetization, flushing, magnetization are repeated, and rinsing each time can loss of energy free nucleic acid；Extraction process needs
Professional is wanted to operate, and heavy workload；Nowadays, the rapid and convenientization extraction for how realizing nucleic acid, improves nucleic acid yield, reduces
Reagent sample consumes, and the loss for reducing extraction process amplifying nucleic acid becomes current urgent problem to be solved.
Invention content
In order to overcome the disadvantages of the above prior art, the purpose of the present invention is to provide one kind based on immiscible phase interface
The five chamber nucleic acid-extracting apparatus and method of power realize the rapid extraction of nucleic acid, reduce the consumption of reagent sample, and reduce and extracted
The loss of journey amplifying nucleic acid.
To achieve the above object, the technical solution that the present invention takes is：
A kind of five chamber nucleic acid-extracting apparatus based on immiscible phase interfacial tension, including sample cavity 1, sample cavity 1 export
It is connected by trapezoidal microchannel 4 with the first immiscible phase chamber 2-1 entrances, the first outlets immiscible phase chamber 2-1 and 5 entrance of washing cavities
It being connected by trapezoidal microchannel 4, the outlet of washing cavities 5 and the second immiscible phase chamber 2-2 entrances are connected by trapezoidal microchannel 4, the
Two outlets immiscible phase chamber 2-2 and elution 3 entrance of chamber are connected by trapezoidal microchannel 4, and the entrance of sample cavity 1 is well 7, the
One immiscible phase chamber 2-1, the second immiscible phase chamber 2-2 are respectively equipped with the first tank filler sleeve 9, the second tank filler sleeve 8, and washing cavities 5 are equipped with
Add and wash fluid apertures 10, elution chamber 3 is equipped with plus elution fluid apertures 11, and the outside of sample cavity 1, immiscible phase chamber 2 or elution chamber 3 is equipped with magnetic
Body 6.
The mouth width of the trapezoidal microchannel 4 is 500 μm, is highly 250 μm.
A kind of extracting method of the five chamber nucleic acid-extracting apparatus based on immiscible phase interfacial tension, includes the following steps：
By biological cell at a temperature of 37 DEG C, cultivate in the medium to converging, carefully by protein enzyme solution release biology
Born of the same parents, and centrifuged, the cell precipitate after centrifugation is collected；Cell precipitate, nanometer magnetic bead are added to lysate buffering
In liquid, it is incubated at room temperature to allow fully cracking, and the nucleic acid reaction knot discharged after making nanometer magnetic bead and biological cell crack
Conjunction forms magnetic bead-nucleic acid complexes；Lysis buffer and magnetic bead-nucleic acid complexes are all injected into sample by well 7
Chamber 1, by elution buffer by adding elution fluid apertures 11 that elution chamber 3 is added, by cleaning solution by adding washing fluid apertures 10 that washing is added
Chamber 5 gives the first immiscible phase chamber 2-1, the second immiscible phase chamber respectively by the first tank filler sleeve 9, the second tank filler sleeve 8 immediately after
2-2 fills immiscible phase；Magnet 6 is rotated in 1 bottom outside of sample chamber so that is discharged after nanometer magnetic bead and biological cell cracking
The abundant reaction bonded of nucleic acid forms magnetic bead-nucleic acid complexes, and magnet 6 is moved by lateral first immiscible phase chamber 2-1 outside sample cavity 1
It is dynamic, then moved from the first immiscible phase chamber 2-1 to washing cavities 5, when magnetic bead-nucleic acid complexes are moved to washing cavities 5, pass through shifting
Liquid pipe blows and beats intracavitary cleaning solution repeatedly, to realize the washing to magnetic bead-nucleic acid complexes, after washing, magnet 6
Again from washing cavities 5 to the second immiscible phase chamber 2-2 movements, finally moved from the second immiscible phase chamber 2-2 to elution chamber 3 so that
Magnetic bead-nucleic acid complexes pass through the first immiscible phase chamber 2-1, washing cavities 5 and the second immiscible phase chamber 2-2 by trapezoidal microchannel 4
It is eluted into elution chamber 3, after elution, nucleic acid is released；By adding in sample cavity 1, elution chamber 3 and washing cavities 5
Enter surfactant to maintain water phase and oil phase interface energy.
The immiscible phase is using silicone oil, olive oil, liquid wax, air or FC-40 oil.
The elution buffer is Tris-HCl (Trisaminomethane) or TE buffer solutions.
The magnet 6 uses permanent magnet or electromagnet.
The cleaning solution is GuHCl (guanidine hydrochloride).
The surfactant be Triton X-100 (Triton X-100), Sarkosyl (flesh aminoacyl) or
Tween-20 (polysorbas20) surfactant.
Gain effect of the present invention is：
(1) it uses the four unmixing interfaces formed at four 4 osculums of trapezoidal microchannel as barrier, and only carries out primary
Washing process, to realize the purifying of nucleic acid.The multiple washing step needed for traditional paramagnetic particle method is eliminated to improve nucleic acid extraction
Efficiency.
(2) washing cavities 5 are so that biological cell cracks the nucleic acid discharged and the abundant reaction bonded of nanometer magnetic bead and washes away portion
Divide impurity, is conducive to the extraction efficiency for improving nucleic acid.
(3) superparamagnetism for utilizing nanometer magnetic bead, realizes the movement of magnetic bead-nucleic acid complexes, compares traditional paramagnetic particle method, do not have
The mechanical movement for having bar magnet reduces the nucleic acid loss in extraction process.
(4) traditional paramagnetic particle method nucleic acid extraction is compared, under the premise of nucleic acid yield is identical, substantially reduces extraction time.
(5) plug structure surrogate response container is isolated using the miniature closing of trapezoidal microchannel 4 connection, reduces examination used
Agent volume, and effectively prevent reagent contamination and evaporation.
(6) apparatus of the present invention structure is simpler, easy to process.
Description of the drawings
Fig. 1 is the vertical view of extraction element of the present invention.
Fig. 2 is the side view of extraction element of the present invention.
Fig. 3 is that nucleic acid extraction of embodiment of the present invention method and two kinds of commercial reagents box amplifications are thin from 5.6 ten thousand breast cancer epitheliums
Three genes (P0, ER α and GR) extracted nucleic acid yield comparison of the mRNA of born of the same parents (MCF-7).
Specific implementation mode
Clear, complete description is carried out to technical scheme of the present invention with reference to the accompanying drawings and examples.But it is as described below,
Only it is present pre-ferred embodiments, limitation in any form, every technology according to the present invention not is done to the present invention
Essence any simple modification made to the above embodiment, equivalent variations and modification, belong to technical solution of the present invention scope.
Referring to Figures 1 and 2, a kind of five chamber nucleic acid-extracting apparatus based on immiscible phase interfacial tension, including sample cavity
1, the outlet of sample cavity 1 and the first immiscible phase chamber 2-1 entrances are connected by trapezoidal microchannel 4, the first outlets immiscible phase chamber 2-1
It is connected by trapezoidal microchannel 4 with 5 entrance of washing cavities, the outlet of washing cavities 5 passes through trapezoidal micro- with the second immiscible phase chamber 2-2 entrances
Channel 4 connects, and the second outlets immiscible phase chamber 2-2 and elution 3 entrance of chamber are connected by trapezoidal microchannel 4, the entrance of sample cavity 1
For well 7, the first immiscible phase chamber 2-1, the second immiscible phase chamber 2-2 are respectively equipped with the first tank filler sleeve 9, the second tank filler sleeve 8,
Washing cavities 5 are equipped with plus washing fluid apertures 10, and elution chamber 3 is equipped with plus elution fluid apertures 11, sample cavity 1, immiscible phase chamber 2 or elution chamber 3
Outside be equipped with magnet 6.The width of 4 osculum of trapezoidal microchannel is 500 μm, is highly 250 μm.
A kind of extracting method of the five chamber nucleic acid-extracting apparatus based on immiscible phase interfacial tension, includes the following steps：
By biological cell at a temperature of 37 DEG C, cultivate in the medium to converging, carefully by protein enzyme solution release biology
Born of the same parents, and centrifuged, the cell precipitate after centrifugation is collected；Cell precipitate, nanometer magnetic bead are added to lysate buffering
In liquid, it is incubated at room temperature to allow fully cracking, and the nucleic acid reaction knot discharged after making nanometer magnetic bead and biological cell crack
Conjunction forms magnetic bead-nucleic acid complexes.Lysis buffer and magnetic bead-nucleic acid complexes are all injected into sample by well 7
Chamber 1, by elution buffer by adding elution fluid apertures 11 that elution chamber 3 is added, by cleaning solution by adding washing fluid apertures 10 that washing is added
Chamber 5 gives the first immiscible phase chamber 2-1, the second immiscible phase chamber respectively by the first tank filler sleeve 9, the second tank filler sleeve 8 immediately after
2-2 fills immiscible phase；Magnet 6 is rotated in 1 bottom outside of sample cavity so that is discharged after nanometer magnetic bead and biological cell cracking
The abundant reaction bonded of nucleic acid forms magnetic bead-nucleic acid complexes, and magnet 6 is moved by lateral first immiscible phase chamber 2-1 outside sample chamber 1
It is dynamic, then moved from the first immiscible phase chamber 2-1 to washing cavities 5, when magnetic bead-nucleic acid complexes are moved to washing cavities 5, pass through shifting
Liquid pipe blows and beats intracavitary cleaning solution repeatedly, to realize the washing to magnetic bead-nucleic acid complexes, after washing, magnet 6
Again from washing cavities 5 to the second immiscible phase chamber 2-2 movements, finally moved from the second immiscible phase chamber 2-2 to elution chamber 3 so that
Magnetic bead-nucleic acid complexes pass through the first immiscible phase chamber 2-1, washing cavities 5 and the second immiscible phase chamber 2-2 by trapezoidal microchannel 4
It is eluted into elution chamber 3, after elution, nucleic acid is released；By adding in sample cavity 1, elution chamber 3 and washing cavities 5
Enter surfactant to maintain water phase and oil phase interface energy.
Extracting method is described in detail with reference to embodiment.
A kind of extracting method of the five chamber nucleic acid-extracting apparatus based on immiscible phase interfacial tension, includes the following steps：
1) under 37 DEG C of temperature conditions, by improved culture medium of the breast cancer epithelial cell culture in polystyrene flask
(DMEM) in, until converging；Cell is discharged using 0.05% trypsin solution, and by being collected by centrifugation, by cell precipitate
It is freezed at -80 DEG C, until carrying out nucleic acid separable programming；
2) by cell precipitate and a concentration of 0.7mg ml-1MagneSil type nanometer magnetic bead solution be added to cracking buffering
In liquid (1%LiDS, 100mM Tris-HCl, 500mM LiCl, 10mM EDTA, 5mM DTT), and it is incubated at room temperature 5 minutes
To allow to crack, and the abundant reaction bonded of nucleic acid discharged after making nanometer magnetic bead and biological cell crack is compound at magnetic bead-nucleic acid
Object；
3) 8.5 μ l lysis buffers and magnetic bead-nucleic acid complexes are added to sample cavity using pipette by well 7
In 1, by 8.5 μ l elution buffers (10mM Tris-HCl) by adding elution fluid apertures 11 to be added to elution chamber 3, by 8.5 μ l washings
Liquid (5M GuHCl) is by adding washing fluid apertures 10 to be added to washing cavities 5；After addition, passed through immediately with the FC-40 oil of 8.5 μ l
First tank filler sleeve 9, the second tank filler sleeve 8 are added separately to the first immiscible phase chamber 2-1, the second immiscible phase chamber 2-2, by sample
Product chamber 1, elution chamber 3 and washing cavities 5 in a concentration of 0.005% surfactant Triton X-100 are added maintain water phase with
Oil phase interface can be in 3-15nN/m；
4) using 6 (12 × 3 × 3mm of magnet for holding neodymium iron boron (NdFeB)3) mobile magnetic bead-nucleic acid complexes are provided
Required magnetic force, magnet 6 are rotated in the bottom outside of sample cavity 1 so that the nucleic acid discharged after nanometer magnetic bead and biological cell cracking
Abundant reaction bonded forms magnetic bead-nucleic acid complexes；Then, hand magnet 6 with the speed less than 5mm/s by the outside of sample chamber 1
It is moved to the first immiscible phase chamber 2-1 movements, then from the first immiscible phase chamber 2-1 to washing cavities 5, when magnetic bead-nucleic acid complexes
When being moved to washing cavities 5, intracavitary cleaning solution is blown and beaten repeatedly by pipette, to realize to magnetic bead-nucleic acid complexes
Washing, after washing, magnet 6 is again from washing cavities 5 to the second immiscible phase chamber 2-2 movement, finally by the second immiscible phase
Chamber 2-2 to elution chamber 3 move so that magnetic bead-nucleic acid complexes by trapezoidal microchannel 4 pass sequentially through the first immiscible phase chamber 2-1,
Washing cavities 5 and the second immiscible phase chamber 2-2 enters elution chamber 3 and is eluted, and after elution, nucleic acid is released；
5) after using the nucleic acid that pipette collects elution buffer and purifying, the concentration of extracting nucleic acid by using
Multiskan TM GO microplate spectrophotometers measure the absorbance of 2nL eluents to assess extracting nucleic acid purity；DNA's
Amplification carries out polymerase chain reaction (PCR) to realize using 96 thermal cyclers of Q-cycler.
To sum up, the entire nucleic acid extraction time is 8.5 minutes, and relatively conventional instrument for extracting nucleic acid, which takes, reduces 54.1%.Table
One is compared for the present embodiment extraction method and using traditional paramagnetic particle method nucleic acid extraction required time, from experimental result as can be seen that originally
The time required to invention extraction method greatly reduces nucleic acid extraction, this has benefited from this method and only eliminates core by primary effectively filtering
The multiple washing step of acid extraction improves nucleic acid extraction efficiency to greatly reduce extraction time.
Table one
Table two is that the present embodiment nucleic acid extraction method is compared with reagent needed for traditional paramagnetic particle method nucleic acid extraction, can from experiment
Go out, extraction method of the present invention greatly reduces required reagent sample volume.
Table two
Fig. 3 is that the present embodiment nucleic acid extraction method comes from 5.6 ten thousand breast cancer epithelial cells with two kinds of commercial reagents box amplifications
(MCF-7) three genes (P0, ER α and GR) extracted nucleic acid yield comparison of mRNA, can be seen that from experiment, present invention extraction
The nucleic acid yield of method extraction is higher.
Claims (9)
1. a kind of five chamber nucleic acid-extracting apparatus based on immiscible phase interfacial tension, including sample cavity (1), sample cavity (1) go out
Mouth and the first immiscible phase chamber (2-1) entrance are connected by trapezoidal microchannel (4), and the first immiscible phase chamber (2-1) is exported and washed
It washs chamber (5) entrance to connect by trapezoidal microchannel (4), washing cavities (5) outlet and second immiscible phase chamber (2-2) entrance pass through ladder
Shape microchannel (4) connects, and the outlet of the second immiscible phase chamber (2-2) and elution chamber (3) entrance are connected by trapezoidal microchannel (4),
The entrance of sample cavity (1) is well (7), and the first immiscible phase chamber (2-1), the second immiscible phase chamber (2-2) are respectively equipped with the
One tank filler sleeve (9), the second tank filler sleeve (8), washing cavities (5) are equipped with plus washing fluid apertures (10), and elution chamber (3) is equipped with plus elution fluid apertures
(11), the outside of sample cavity (1), immiscible phase chamber (2) or elution chamber (3) is equipped with magnet (6).
2. five chambers nucleic acid-extracting apparatus according to claim 1, it is characterised in that：The trapezoidal microchannel (4)
Width is 500 μm, is highly 250 μm.
3. the extracting method of five chambers nucleic acid-extracting apparatus according to claim 1, which is characterized in that including following step
Suddenly：
By biological cell at a temperature of 37 DEG C, is cultivated in the medium to converging, biological cell is discharged by protein enzyme solution, and
It is centrifuged, the cell precipitate after centrifugation is collected；Cell precipitate, nanometer magnetic bead are added in lysate buffer solution,
It is incubated at room temperature to allow fully cracking, and the nucleic acid reaction discharged after making nanometer magnetic bead and biological cell crack is combined and formed
Magnetic bead-nucleic acid complexes；Lysis buffer and magnetic bead-nucleic acid complexes are all injected into sample cavity by well (7)
(1), by elution buffer by adding elution fluid apertures (11) that elution chamber (3) is added, by cleaning solution by adding detergent hole (10) to add
Enter washing cavities, gives the first immiscible phase chamber (2-1), second not by the first tank filler sleeve (9), the second tank filler sleeve (8) immediately after
Miscible phase chamber (2-2) fills immiscible phase；Magnet (6) is rotated in sample chamber (1) bottom outside so that nanometer magnetic bead and biological sample
The abundant reaction bonded of product forms magnetic bead-nucleic acid complexes, by magnet (6) by sample chamber (1) lateral first immiscible phase chamber (2- outside
1) mobile then mobile from the first immiscible phase chamber (2-1) to washing cavities (5), then from washing cavities (5) to the second immiscible phase chamber
(2-2) is mobile, finally mobile from the second immiscible phase chamber (2-2) to elution chamber (3) so that magnetic bead-nucleic acid complexes are by trapezoidal
Microchannel (4) enters elution chamber (3) by the first immiscible phase chamber (2-1), washing cavities (5) and the second immiscible phase chamber (2-2)
It is eluted, after elution, nucleic acid is released, by the way that surface is added in sample cavity (1), elution chamber (3) and washing cavities (5)
Activating agent maintains water phase and oil phase interface energy.
4. extracting method according to claim 3, it is characterised in that：The immiscible phase is using silicone oil, olive oil, liquid
Body wax, air or FC-40 oil.
5. extracting method according to claim 3, it is characterised in that：The elution buffer is Tris-HCl (three ammonia
Methylmethane) or TE buffer solutions.
6. extracting method according to claim 3, it is characterised in that：The magnet (6) uses permanent magnet or electromagnet.
7. extracting method according to claim 3, it is characterised in that：The cleaning solution is GuHCl (guanidine hydrochloride).
8. extracting method according to claim 3, it is characterised in that：The surfactant is Triton X-100
(Triton X-100), Sarkosyl (flesh aminoacyl) or Tween-20 (polysorbas20) surfactant.
9. extracting method according to claim 3, which is characterized in that include the following steps：
1) under 37 DEG C of temperature conditions, by improved culture medium (DMEM) of the breast cancer epithelial cell culture in polystyrene flask
In, until converging；Cell is discharged using 0.05% trypsin solution, and by being collected by centrifugation, by cell precipitate at -80 DEG C
Lower freezing, until carrying out nucleic acid separable programming；
2) by cell precipitate and a concentration of 0.7mg ml-1MagneSil type nanometer magnetic bead solution be added to lysis buffer
In, lysis buffer 1%LiDS, 100mM Tris-HCl, 500mM LiCl, 10mM EDTA, 5mM DTT, and at room temperature
5 minutes are incubated to allow to crack, and the abundant reaction bonded of nucleic acid discharged after making nanometer magnetic bead and biological cell crack is at magnetic bead-
Nucleic acid complexes；
3) 8.5 μ l lysis buffers and magnetic bead-nucleic acid complexes are added to sample cavity using pipette by well (7)
(1) in, by 8.5 μ l elution buffers by adding elution fluid apertures (11) to be added to elution chamber (3), elution buffer 10mM
Tris-HCl, by 8.5 μ l cleaning solutions by adding washing fluid apertures (10) to be added to washing cavities (5), cleaning solution is 5M GuHCl；Addition
After, it is added separately to first by the first tank filler sleeve (9), the second tank filler sleeve (8) with the FC-40 oil of 8.5 μ l immediately and not mixed
Molten phase chamber (2-1), the second immiscible phase chamber (2-2) are dense by being added in sample cavity (1), elution chamber (3) and washing cavities (5)
It spends and maintains water phase and the oil phase interface to be in 3-15nN/m for 0.005% surfactant Triton X-100；
4) magnetic force needed for mobile magnetic bead-nucleic acid complexes, magnet (6) are provided using the magnet (6) of hand-held neodymium iron boron (NdFeB)
Specification is 12 × 3 × 3mm3, bottom outside rotation of the magnet (6) in sample cavity (1) so that nanometer magnetic bead is cracked with biological cell
The abundant reaction bonded of nucleic acid discharged afterwards forms magnetic bead-nucleic acid complexes；Then, hand magnet (6) is with the speed less than 5mm/s
By sample chamber (1), lateral first immiscible phase chamber (2-1) is mobile outside, then is moved from the first immiscible phase chamber (2-1) to washing cavities (5)
It is dynamic, when magnetic bead-nucleic acid complexes are moved to washing cavities (5), intracavitary cleaning solution is blown and beaten repeatedly by pipette, to
Realize washing to magnetic bead-nucleic acid complexes, after washing, magnet (6) is again from washing cavities (5) to the second immiscible phase chamber
(2-2) is mobile, finally mobile from the second immiscible phase chamber (2-2) to elution chamber (3) so that magnetic bead-nucleic acid complexes are by trapezoidal
Microchannel (4) passes sequentially through the first immiscible phase chamber (2-1), washing cavities (5) and the second immiscible phase chamber (2-2) and enters elution chamber
(3) it is eluted, after elution, nucleic acid is released；
5) after using pipette to collect elution buffer and the nucleic acid of purifying, the concentration of extracting nucleic acid is by using Multiskan
TM GO microplate spectrophotometers measure the absorbance of 2nL eluents to assess extracting nucleic acid purity；The amplification of DNA uses Q-
96 thermal cyclers of cycler carry out polymerase chain reaction (PCR) to realize.
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| CN111057638A (en) * | 2018-10-17 | 2020-04-24 | 北京致雨生物科技有限公司 | Sample processing device and method, and digital PCR system including the same |
| CN112266841A (en) * | 2020-10-23 | 2021-01-26 | 东南大学 | Biological sample processing chip device and processing method |
| CN112442443A (en) * | 2020-06-11 | 2021-03-05 | 杭州天微基因科技有限公司 | Nucleic acid extraction and amplification integrated detection method and detection connecting pipe |
| CN112538425A (en) * | 2020-10-23 | 2021-03-23 | 北京理工大学 | On-chip nucleic acid amplification detection system and method based on micro-fluidic chip |
| CN112877405A (en) * | 2021-02-22 | 2021-06-01 | 新格元(南京)生物科技有限公司 | Single cell lysis solution and application thereof |
| CN113337398A (en) * | 2021-05-31 | 2021-09-03 | 深圳市博德致远生物技术有限公司 | Micro-fluidic chip |
| CN114381368A (en) * | 2021-12-31 | 2022-04-22 | 北京理工大学 | System and method for automatically separating biological analytes by using gas-liquid interface |
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