CN205774469U - One-step method prepares the device of fragmentation DNA - Google Patents
One-step method prepares the device of fragmentation DNA Download PDFInfo
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- CN205774469U CN205774469U CN201620501344.5U CN201620501344U CN205774469U CN 205774469 U CN205774469 U CN 205774469U CN 201620501344 U CN201620501344 U CN 201620501344U CN 205774469 U CN205774469 U CN 205774469U
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- centrifuge tube
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Abstract
This utility model relates to a new generation's high-throughput sequencing library and builds field of sample preparation, especially relates to the device that a kind of one-step method prepares fragmentation DNA.Ultrasonic cleaner is provided with cover plate, and cover plate lower surface arranges the baffle plate of symmetry, and cover plate is connected by spring with pressing plate, and the centrifuge tube fixed plate with centrifuge tube is connected with baffle plate grafting.This utility model device acquisition fragmentation DNA efficient, simple and direct, quick, gained fragmentation DNA can may be used for a new generation's high-throughput sequencing library structure.
Description
Technical field
This utility model relates to a new generation's high-throughput sequencing library and builds field of sample preparation, especially
Relate to the device that a kind of one-step method prepares fragmentation DNA.
Background technology
Can construct high-quality library be the key obtaining high-quality sequencing data, and obtains
The DNA fragmentation of suitable size is then premise and the basis building high-quality library.Conventional DNA
The method of fragmentation mainly has two kinds: one is enzyme cutting method, mainly utilizes specific enzyme identification
Restriction enzyme site on genome sequence is by genomic fragment；Another kind is Physical, mainly profit
By the way of Mechanical Crushing, genome is interrupted at random, such as atomization, mechanical shearing, ultrasonic beat
Break, most common of which use sonicator by genomic DNA fragment exactly.
The model of sonicator and kind are all a lot, and supersonic frequency and power and price are also
The most different.The sonicator being currently used for high-flux sequence DNA sample broken is main
Have: contactless fully-automatic ultrasonic crushes instrument Bioruptor and automatic focused sound waves sample process
Instrument Covaris, but their volume is the most relatively large, takies laboratory relatively multiple resource, and
Expensive.
But, now all of sonicator is all directed to DNA sample and operates,
Need to complete by conventional DNA extraction method the extraction of genomic DNA before this, pass through
56 DEG C of water-baths of 15min-2h and 70 DEG C of water-bath cracking of 10min are organized or cell, cross post,
Eluting, then could eluting genomic DNA, this process takes around 3 hours and completes.
In view of the above, now be badly in need of one can cost-effective, improve the DNA extraction time and
Simplify complicated operating procedure, and the library structure of different length Insert Fragment size can also be met
The one-step method building demand prepares the device of fragmentation DNA.
Utility model content
This utility model purpose there is provided the device that a kind of one-step method prepares fragmentation DNA.
In order to realize above-mentioned technical purpose, technical solutions of the utility model are:
One-step method prepares the device of fragmentation DNA, and ultrasonic cleaner 7 is provided with cover plate 1,
Cover plate 1 lower surface arranges the baffle plate 3 of symmetry, and cover plate 1 is connected by spring 2 with pressing plate 4,
Centrifuge tube fixed plate 5 with centrifuge tube 6 is connected with baffle plate 3 grafting.
Described cover plate 1 lower surface arranges multiple spring 2, is connected with pressing plate 4 by multiple springs 2；
Pressing plate 4 is squeezed in above the centrifuge tube fixed plate 5 with centrifuge tube 6.The effect of pressing plate is handle
Centrifuge tube lid in fixed plate compresses, and prevents sample from leaking outside, and centrifuge tube fixed plate is front and back to live
Body, can change different size of centrifuge tube.
Described cover plate 1 lower surface arranges two " L " type baffle plates 3 of symmetry, with centrifuge tube 6
Centrifuge tube fixed plate 5 be connected with the minor face grafting of " L " type baffle plate 3.
Multiple hole housing centrifuge tube 6 is set in described fixed plate 5.
Described employing said apparatus concrete fragmentation DNA preparation mainly comprises the steps of
1) sample process: be added to 1.5ml centrifuge tube 6 by needing broken a certain amount of sample
In, 10,000g are centrifuged 1min, abandon supernatant；
2) in above-mentioned centrifuge tube 6,80-100ul lysate is added；
3) device for opening cover plate 1, adds frozen water mixed liquor to device ultrasonic cleaner 7
In rinse bath, being allowed to be maintained at 0 DEG C, liquid level is advisable with liquid level in somewhat not having centrifuge tube；
4) above-mentioned centrifuge tube 6 is inserted in fixed plate 5, opens instrument switch, according to institute
Need the size of DNA fragmentation, different ultrasonic times is set；
5) close the lid this device of operation；
6) ultrasonic end, takes out centrifuge tube, extracts the DNA of fragmentation.
Described sample, its source can be plant, animal or microbial cell, it is also possible to is
The cell suspension that tissue samples is made after grinding.
Ibid aforesaid way acquisition fragmentation DNA may be used for new-generation sequencing library construction, and
It is applied to machine order-checking on high flux.
This utility model is had an advantage in that
Simple and the cheap ultrasonic cleaner of this utility model device binding operation and specific
Fixing device, interrupts ultrasound wave and is combined in one with the cleavage step in the DNA extraction of cell sample
Step completes, time-consuming and need Pintsch process tissue or thin in the conventional DNA extraction kit of omission
The step of born of the same parents, directly cross post, scrubbed after can elute the highly purified fragmentation of high-quality
DNA, the DNA of obtained fragmentation are used directly for next step library construction.
Accompanying drawing explanation
Fig. 1 is apparatus structure schematic diagram of the present utility model；Wherein, 1. cover plate；2. spring；
3. spring shield；4. pressing plate；5. centrifuge tube fixed plate；6. centrifuge tube；7. ultrasonic cleaner.
Fig. 2 is the electricity using apparatus of the present invention to extract mouth epithelial cells sample DNA broken results
Swimming figure.
Fig. 3 is that the fragment using biological big analyser to detect mouth epithelial cells fragmentation DNA is big
Little scattergram.
Detailed description of the invention
This utility model device can acquisition fragmentation DNA efficient, simple and direct, quick, obtained sheet
Sectionization DNA may be used for a new generation's high-throughput sequencing library and builds.
Describe this utility model below with reference to the accompanying drawings and in conjunction with the embodiments in detail.Following enforcement
Method described in example is conventional method if no special instructions.
Embodiment 1:
Seeing the device that Fig. 1 one-step method prepares fragmentation DNA, ultrasonic cleaner 7 is provided with
Cover plate 1, cover plate 1 lower surface arranges the baffle plate 3 of symmetry, and cover plate 1 and pressing plate 4 pass through spring
2 are connected, and the centrifuge tube fixed plate 5 with centrifuge tube 6 is connected with baffle plate 3 grafting.
Described cover plate 1 lower surface arranges multiple spring 2, is connected with pressing plate 4 by multiple springs 2；
Pressing plate 4 is squeezed in above the centrifuge tube fixed plate 5 with centrifuge tube 6.The effect of pressing plate is handle
Centrifuge tube lid in fixed plate compresses, and prevents sample from leaking outside, and centrifuge tube fixed plate is front and back to live
Body, can change different size of centrifuge tube.
Described cover plate 1 lower surface arranges two " L " type baffle plates 3 of symmetry, with centrifuge tube 6
Centrifuge tube fixed plate 5 be connected with the minor face grafting of " L " type baffle plate 3.
Multiple hole housing centrifuge tube 6 is set in described fixed plate 5.
Embodiment 2
Utilize embodiment 1 device system to mouth epithelial cells 250-550bp fragmentation DNA
Standby:
1) broken mouth epithelial cells sample will be needed to be added in 1.5ml centrifuge tube 6,
10,000g are centrifuged 1min, abandon supernatant, the general 10ul of sample pellet amount；
2) adding the existing lysate of 90ul in above-mentioned centrifuge tube, cumulative volume is 100ul；
3) device for opening cover plate 1, adds frozen water mixed liquor to device ultrasonic cleaner 7
In rinse bath, being allowed to be maintained at 0 DEG C, liquid level is advisable with liquid level in somewhat not having centrifuge tube；
4) above-mentioned centrifuge tube 6 is inserted in fixed plate 5, opens instrument switch, according to institute
Needing the size of DNA fragmentation, arranging ultrasonic time is 30s；
5) close the lid this device of operation, and centrifuge tube is taken out in ultrasonic end；
6) use QIAamp DNA mini kit to extract genomic DNA, eliminate this DNA
Extract the step of Pintsch process cell in test kit, directly cross post, wash and elute fragmentation
DNA；
7) said extracted DNA out is detected its concentration through microplate reader Tecan-100
44.5ng/ul, OD value is 1.83；
8) take out appropriate DNA to verify through 1% sepharose electrophoresis, obtained 250-550bp's
Fragment, as shown in Figure 2.
9) above-mentioned DNA sample is detected with biomacromolecule analyser Labchip further,
Result is consistent with sepharose electrophoresis, broken abundant, obtains the band of even dispersion, such as accompanying drawing 3.
The foregoing is only preferred embodiment of the present utility model, be not limited to this reality
With novel, for a person skilled in the art, this utility model can have various change and
Change.All within spirit of the present utility model and principle, any amendment, the equivalent made are replaced
Change, improvement etc., within should be included in protection domain of the present utility model.
Claims (4)
1. an one-step method prepares the device of fragmentation DNA, it is characterised in that: ultrasound wave is clear
Washing device (7) and be provided with cover plate (1), cover plate (1) lower surface arranges the baffle plate (3) of symmetry,
Cover plate (1) is connected by spring (2) with pressing plate (4), being centrifuged with centrifuge tube (6)
Pipe fixed plate (5) is connected with baffle plate (3) grafting.
2. the one-step method as described in claim 1 prepares the device of fragmentation DNA, and its feature exists
In: described cover plate (1) lower surface arranges multiple spring (2), by multiple springs (2)
It is connected with pressing plate (4)；
Pressing plate (4) is squeezed in centrifuge tube fixed plate (5) top with centrifuge tube (6).
3. the one-step method as described in claim 1 prepares the device of fragmentation DNA, its feature
It is: described cover plate (1) lower surface arranges two " L " type baffle plates (3) of symmetry, band
There are the centrifuge tube fixed plate (5) of centrifuge tube (6) and the minor face grafting of " L " type baffle plate (3)
It is connected.
4. the one-step method as described in claim 1 prepares the device of fragmentation DNA, its feature
It is: multiple hole housing centrifuge tube (6) is set in described fixed plate (5).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201620501344.5U CN205774469U (en) | 2016-05-27 | 2016-05-27 | One-step method prepares the device of fragmentation DNA |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201620501344.5U CN205774469U (en) | 2016-05-27 | 2016-05-27 | One-step method prepares the device of fragmentation DNA |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN205774469U true CN205774469U (en) | 2016-12-07 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201620501344.5U Expired - Fee Related CN205774469U (en) | 2016-05-27 | 2016-05-27 | One-step method prepares the device of fragmentation DNA |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109266545A (en) * | 2018-09-29 | 2019-01-25 | 宁波新芝生物科技股份有限公司 | A kind of ultrasonic wave DNA interrupts instrument |
-
2016
- 2016-05-27 CN CN201620501344.5U patent/CN205774469U/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109266545A (en) * | 2018-09-29 | 2019-01-25 | 宁波新芝生物科技股份有限公司 | A kind of ultrasonic wave DNA interrupts instrument |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20161207 Termination date: 20180527 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |