CN209010540U - A kind of dissociative DNA extracts kit - Google Patents
A kind of dissociative DNA extracts kit Download PDFInfo
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- CN209010540U CN209010540U CN201820843725.0U CN201820843725U CN209010540U CN 209010540 U CN209010540 U CN 209010540U CN 201820843725 U CN201820843725 U CN 201820843725U CN 209010540 U CN209010540 U CN 209010540U
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- test tube
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- dissociative dna
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Abstract
The utility model discloses a kind of dissociative DNA extracts kits, comprising: box body, test tube flat, controller, fixed frame, temperature sensor, takes object lid, oscillation device, heating device, protrusion, movable plate, jacking block, sliding rail, support unit at box cover；The utility model realizes that test tube flat slides up and down in the cavity of box body, test tube flat can directly be sent out in the cavity of fixing layer together with test tube, take test tube convenient for operator using the position for adjusting movable plate, easy to operate.It is provided with heating device and oscillation device, temperature and frequency of oscillation is controlled respectively, ensure that the high efficiency of kit.In addition, agent formulations are simple, synergistic effect is strong, and stability is good, and product purity is high, the quick separating DNA suitable for the cell-free body fluid such as serum or blood plasma.
Description
Technical field
The utility model belongs to field of medical immunology more particularly to a kind of dissociative DNA extracts kit.
Background technique
DNA is the carrier of hereditary information, is one of main molecular biology research object, with molecular biology research
Deepen continuously and the fast development of gene sequencing technology, time and cost rapid decrease needed for gene sequencing, in gene water
The basic fundamental means that diagnosis has become clinical diagnosis are carried out to disease on flat, the accurate medical treatment based on gene sequencing is just
It is increasingly becoming the important way in the fields such as major disease diagnosis, personalized medicine, fetus genetic disease credit analysis and individual identification.
After normal cell, tumour cell or fetal tissue's cell division, the DNA in cell can be discharged into human body fluid,
It is free in except cell, wherein this part DNA for entering blood is referred to as dissociative DNA in blood, also referred to as plasma DNA,
That is cell free DNA, abbreviation cfDNA.CfDNA fragment length is concentrated mainly between 100~240bp, it is most of 160~
Between 200bp.Dissociative DNA is a kind of extracellular dna of cell-free state, mainly by single-stranded or double-stranded DNA and it is single-stranded with it is double
The mixture of chain DNA forms, and exists in the form of mono- protein complex of DNA or two kinds of dissociative DNA.Healthy human peripheral blood there is also
CfDNA, but the peripheral blood cfDNA content of tumor patient has increasing for conspicuousness compared with healthy human peripheral blood cfDNA, and swollen
There are gene mutation, recombination, missings etc. in tumor peripheral blood in patients DNA.Blood of cancer patients dissociative DNA is mainly tumour cell
Necrosis or the tumour cell of apoptosis, the cracking of micrometastasis stove or circulating tumor cell and molecular marker for increased proliferation discharged.
It is patient's primary tissue sample that oncogene, which detects most direct object, but those were not carried out with the swollen of operation
Tumor patient, tumor specimen are not easy to obtain, though and the technology of Biopsy is more mature, patient acceptance is relatively low, especially
Its patient that those with many places transfer have occurred.Even if there are samples before, but often even protect several years ago a few months ago
The local sample deposited, dynamic and heterogeneity in view of Oncogenome, the information that they reflect has been out-of-date or generation
The information of table is not complete.Clinical research shows that cfDNA can reflect the tumor load, grade malignancy, transfer energy of patient's entirety well
Power and real-time gene mutation information have preferable consistency with the gene information of tumor tissues.What blood plasma cfDNA was extracted
Quality is the guarantee to DNA mutation detection, and the method for extracting cfDNA at present mainly has phenol chloroform isoamyl alcohol method, column formulation, magnetic bead
Method.Traditional phenol chloroform isoamyl alcohol method is to extract DNA all in blood plasma to the extraction of blood plasma cfDNA, cannot be by blood plasma
Middle size segment separation, extraction efficiency is low, its efficiency need to be improved by other means.Column formulation extraction efficiency is high, but can not
Different size of DNA fragmentation is separated.Paramagnetic particle method is the functional group absorption nucleic acid using nanoscale magnetic bead bead surface, is utilized
The volume ratio of magnetic bead and solution sorts different size DNA, the higher rate of recovery not only can be obtained, but also can be to cfDNA
It is sorted.Research prompt tumor patient periphery dissociative DNA segment is small compared with normal person's dissociative DNA, in colorectal cancer patients blood plasma
80% or more dissociative DNA segment is respectively less than 145bp, this prompt tumour cell release dissociative DNA may small fragment ratio compared with
Greatly, therefore by tumor patient periphery dissociative DNA size segment it separates, to being based further on peripheral circulation tumor cell DNA
Monitoring oncogene information is of great significance.
Utility model content
In view of the shortcomings of the prior art, the utility model provides a kind of dissociative DNA extracts kits, comprising: box body, box
Lid, test tube flat, controller, fixed frame, temperature sensor, take object lid, oscillation device, heating device, protrusion, movable plate, jacking block,
Sliding rail, support unit.
The upper surface of the box body is provided with box cover, and is provided on the box cover corresponding with test tube position
Taking mouth, and be screwed in taking mouth and take object lid.Fixed frame is equipped in the box body, opened up on the fixed frame there are four
The inner wall of transparent cavity, each cavity is equipped with four protrusions arranged vertically；It is equipped in each cavity for putting
The test tube flat of test tube is set, the test tube flat is equipped with several blind holes for holding test tubes；And the test tube flat side
It is provided with oscillation device, wherein the oscillation device and controller is electrically connected.The test tube flat bottom is provided with heating
Device is additionally provided with temperature sensor in the box body wherein the heating device and controller is electrically connected.Described
Movable plate is equipped with positioned at the lower section of fixed frame in box body, and the upper surface of the movable plate sets that there are two can jack up test tube flat
Jacking block.The width of the movable plate and the width of box body are equal, and the length of movable plate is greater than the length of box body, the box
Body, which is located at below fixed frame, is symmetrically arranged with two sliding rails on the inner sidewall of corresponding movable plate long side.In addition, the test tube flat
Bottom is additionally provided with support unit.Test tube in blind hole be placed with respectively magnetic bead pretreatment fluid, cell cracking combination liquid, cleaning solution I,
Cleaning solution II, eluent.
The groove with male cooperation is formed on the side wall of the test tube flat.
The heating device is heating tube.
The signal end of the temperature sensor is connect with controller.
Its described movable plate is cuboid.
The height of the jacking block is less than movable plate at a distance from heating device bottom.
The utility model has the advantages that
A kind of dissociative DNA extracts kit of the utility model realizes test tube flat in box body using the position for adjusting movable plate
Cavity in slide up and down, test tube flat can directly be sent out in the cavity of fixing layer together with test tube, be taken convenient for operator
Test tube, it is easy to operate.It is provided with heating device and oscillation device, temperature and frequency of oscillation is controlled respectively, ensure that kit
High efficiency.In addition, agent formulations are simple, synergistic effect is strong, and stability is good, and product purity is high, is suitable for the nothings such as serum or blood plasma
Quick separating DNA in cell body fluid.
Detailed description of the invention:
Fig. 1 is a kind of schematic perspective view of dissociative DNA extracts kit of the utility model；
Fig. 2 is the structural schematic diagram of the utility model box cover；
Fig. 3 is a kind of structural schematic diagram of dissociative DNA extracts kit of the utility model；
Fig. 4 is the flow chart that the utility model dissociative DNA extracts.
Specific embodiment
Below by drawings and examples, the technical solution of the utility model is described in further detail.
As shown in Figures 1 to 4 be a kind of dissociative DNA extracts kit, comprising: box body 1, box cover 2, test tube flat 3,
Controller 4, temperature sensor 7, takes object lid 8, oscillation device 9, heating device 10, raised 11, movable plate 16, jacking block at fixed frame 6
17, sliding rail 18, support unit 19.
The upper surface of the box body 1 is provided with box cover 2, and is provided on the box cover 2 opposite with test tube position
The taking mouth answered, and be screwed in taking mouth and take object lid 8.It is equipped with fixed frame 6 in the box body 1, is opened on the fixed frame 6
If the inner wall of each cavity is equipped with four protrusions 11 arranged vertically there are four transparent cavity；In each cavity
Equipped with the test tube flat 3 for holding test tubes, the groove with 11 cooperation of protrusion is formed on the side wall of the test tube flat 3, it is described
Test tube flat 3 be equipped with several blind holes for holding test tubes；And 3 side of test tube flat is provided with oscillation device 9,
Described in oscillation device 9 and controller 4 be electrically connected, pass through controller 4 control oscillation device 9.3 bottom of test tube flat
It is provided with heating device 10, wherein the heating device 10 and controller 4 is electrically connected, is additionally provided in the box body 1
Temperature sensor 7, and connect in the signal end of the temperature sensor 7 with controller 4.It is controlled by controller 4 described
Thermal 10 temperature, and the temperature that will test by temperature sensor 7 is transmitted back to controller 4 by signal.Described
It is equipped with movable plate 16 positioned at the lower section of fixed frame 3 in box body 1, wherein the movable plate 16 is cuboid, the movable plate
16 upper surface sets that there are two the jacking blocks 17 that can jack up test tube flat 3.The height of the jacking block 17 be less than movable plate 16 with
The width of the distance of 10 bottom of heating device, the movable plate 16 is equal with the width of box body 1, and the length of movable plate 16 is greater than
The length of box body 1, the box body 1, which is located on the inner sidewall for corresponding to 16 long side of movable plate below fixed frame 6, is symmetrically arranged with two
Sliding rail 18, the movable plate 16 can be slided along sliding rail 18.In addition, the bottom of the test tube flat 3 is additionally provided with support unit
19.Test tube in blind hole is placed with magnetic bead pretreatment fluid, cell cracking combination liquid, cleaning solution I, cleaning solution II, eluent respectively.
The magnetic bead pretreatment fluid includes: the nonpolar amino acid of 500ppm and the oligofructose of 1000ppm, wherein non-
Polar amino acid is made of leucine and valine by the weight ratio of 1:0.5.
The cell cracking combination liquid specifically includes that guanidinium isothiocyanate, sodium citrate, detergent, isopropanol.Wherein go
Dirty agent is Triton 100.Under the conditions of high salt conditions and suitable concentration isopropanol, the dissociative DNA in blood plasma can be efficiently
In conjunction with magnetic bead, while removing the impurity contained in the blood plasma such as part albumen, polysaccharide, blood lipid；The cell cracking combination liquid
Middle guanidinium isothiocyanate, sodium citrate, Triton 100, isopropanol content be respectively 50% (W/V), 2-3% (W/V), 10%
(W/V), 2% (W/V).
The cleaning solution I specifically includes that Licl, ethyl alcohol, wherein the content of Licl and ethyl alcohol is in the cleaning solution
5mol/L, 40% (W/V).
The cleaning solution II includes: 75% ethyl alcohol；
The eluent includes: 15mmol/LTris-Hcl and 1mmol/L EDTA, pH 8.0；
Implementation column dissociative DNA extracting method
Step 1: the base magnetic bead is handled by the magnetic bead pretreatment fluid, and the processing time is 5min, standby
With；
Step 2: taking centrifuge tube, and the blood plasma or serum of fresh separated is added, and cell cracking combination liquid is added, turns upside down
It mixes, above-mentioned carboxylated magnetic bead after pretreatment is added, be vortexed and mix, 65 DEG C of water-bath 5min；
Step 3: being cooled to room temperature, places 10 minutes.
Step 4: carrying out Beads enrichment, removes supernatant, retains magnetic bead
Step 5: cleaning solution I being added into centrifuge tube, is vortexed and mixes, and centrifugation discards supernatant；
Step 6: cleaning solution II being added into centrifuge tube, is vortexed and mixes, and centrifugation discards supernatant；
Step 7: uncapping, drying at room temperature 10min；
Step 8: being added eluent, is vortexed and mixes, carries out heating elution, makes magnetic bead dispersion with solution, is stored at room temperature
5min；Brief centrifugation, Aspirate supernatant body, -20 DEG C or -80 DEG C preservations, this liquid are the blood plasma or free serum extracted
DNA。
In conclusion the content of the utility model is not limited in the above embodiments, those skilled in the art can
To propose other embodiments within technological guidance's thought of the invention, but these embodiments are included in the scope of the present invention
Within.
Claims (6)
1. a kind of dissociative DNA extracts kit characterized by comprising box body, box cover, test tube flat, controller, fixed frame, temperature
Degree sensor takes object lid, oscillation device, heating device, protrusion, movable plate, jacking block, sliding rail, support unit；
The upper surface of the box body is provided with box cover, and is provided on the box cover and corresponding with test tube position takes object
Mouthful, and be screwed in taking mouth and take object lid；It is equipped with fixed frame in the box body, is opened up on the fixed frame there are four above and below
The inner wall of penetrating cavity, each cavity is equipped with four protrusions arranged vertically；It is equipped in each cavity for placing examination
The test tube flat of pipe, the test tube flat are equipped with several blind holes for holding test tubes；And the test tube flat side setting
There is oscillation device, wherein the oscillation device and controller is electrically connected；The test tube flat bottom is provided with heating device,
Wherein the heating device and controller are electrically connected, and are additionally provided with temperature sensor in the box body；The box body
The interior lower section positioned at fixed frame be equipped with movable plate, the upper surface of the movable plate set there are two can by test tube flat jack up top
Block；The width of the movable plate and the width of box body are equal, and the length of movable plate is greater than the length of box body, the box body position
Two sliding rails are symmetrically arranged on the inner sidewall for corresponding to movable plate long side below fixed frame；In addition, the bottom of the test tube flat
It is additionally provided with support unit；Test tube in blind hole is placed with magnetic bead pretreatment fluid, cell cracking combination liquid, cleaning solution I, washing respectively
Liquid II, eluent.
2. a kind of dissociative DNA extracts kit according to claim 1, it is characterised in that: the side wall of the test tube flat
On form groove with male cooperation.
3. a kind of dissociative DNA extracts kit according to claim 1, it is characterised in that: the heating device is to add
Heat pipe.
4. a kind of dissociative DNA extracts kit according to claim 1, it is characterised in that: the temperature sensor
Signal end is connect with controller.
5. a kind of dissociative DNA extracts kit according to claim 1, it is characterised in that: its described movable plate is length
Cube.
6. a kind of dissociative DNA extracts kit according to claim 1, it is characterised in that: the height of the jacking block is small
In movable plate at a distance from heating device bottom.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201820843725.0U CN209010540U (en) | 2018-06-01 | 2018-06-01 | A kind of dissociative DNA extracts kit |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201820843725.0U CN209010540U (en) | 2018-06-01 | 2018-06-01 | A kind of dissociative DNA extracts kit |
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| CN209010540U true CN209010540U (en) | 2019-06-21 |
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| CN201820843725.0U Active CN209010540U (en) | 2018-06-01 | 2018-06-01 | A kind of dissociative DNA extracts kit |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111471577A (en) * | 2020-05-22 | 2020-07-31 | 杭州同创医学检验实验室有限公司 | Gene dissociation DNA extraction system |
-
2018
- 2018-06-01 CN CN201820843725.0U patent/CN209010540U/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111471577A (en) * | 2020-05-22 | 2020-07-31 | 杭州同创医学检验实验室有限公司 | Gene dissociation DNA extraction system |
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