CN206736220U - A kind of device that DNA is crushed suitable for high-flux sequence - Google Patents
A kind of device that DNA is crushed suitable for high-flux sequence Download PDFInfo
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- CN206736220U CN206736220U CN201720386522.9U CN201720386522U CN206736220U CN 206736220 U CN206736220 U CN 206736220U CN 201720386522 U CN201720386522 U CN 201720386522U CN 206736220 U CN206736220 U CN 206736220U
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- 238000005406 washing Methods 0.000 claims abstract description 23
- 239000011435 rock Substances 0.000 claims abstract description 13
- 239000000463 material Substances 0.000 claims abstract description 8
- 230000004044 response Effects 0.000 claims description 8
- 239000006260 foam Substances 0.000 claims description 3
- 239000000725 suspension Substances 0.000 claims description 2
- 238000013467 fragmentation Methods 0.000 abstract description 6
- 238000006062 fragmentation reaction Methods 0.000 abstract description 6
- 238000004519 manufacturing process Methods 0.000 abstract 1
- 238000000034 method Methods 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 108020004707 nucleic acids Proteins 0.000 description 8
- 150000007523 nucleic acids Chemical class 0.000 description 8
- 102000039446 nucleic acids Human genes 0.000 description 8
- 230000000694 effects Effects 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 239000010813 municipal solid waste Substances 0.000 description 5
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- 238000005119 centrifugation Methods 0.000 description 4
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- 238000005516 engineering process Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 238000012163 sequencing technique Methods 0.000 description 4
- 238000002604 ultrasonography Methods 0.000 description 4
- 229920000936 Agarose Polymers 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000001502 gel electrophoresis Methods 0.000 description 3
- 239000005457 ice water Substances 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 230000009471 action Effects 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 230000004907 flux Effects 0.000 description 2
- 238000012165 high-throughput sequencing Methods 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 230000007035 DNA breakage Effects 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
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Abstract
The utility model provides a kind of device of the broken DNA suitable for high-flux sequence, belongs to DNA fragmentation preparing technical field.The device includes square DNA sample fixed plate (1) and ultrasonic washing instrument, offered in the plate face of the DNA sample fixed plate and put sample hole (3), described put is provided with heat emission hole (4) around sample hole (3), one jiao of the DNA sample fixed plate (1) is set to positioning and smears angle (2), and groove on the rocks (6) is further opened with described DNA sample fixed plate (1).Present apparatus materials are easy, simple for production, easy to use, work well.
Description
Technical field
Gene high throughput sequencing technologies field is the utility model is related to, in particular it relates to which a kind of be applied to high-flux sequence
In crush DNA device.
Background technology
In two generations and three generations's sequencing technologies, the sequencing library for building high quality is the key factor of high-flux sequence.Survey
The quality in preface storehouse directly determines that the quality of data of output is sequenced, and then has greatest association to data analysis.With high pass
The fast development of sequencing technologies is measured, major Reagent Company, high-flux sequence is in zoology, botany, microbiology, agricultural, doctor
Etc. every field has obtained great application.
In the beginning of sequencing library structure, first fragmentation is carried out to DNA sample.At present, the method for DNA fragmentation has more
Kind, such as enzyme cutting method, adapted local cosine transform ultrasonic wave (AFA, Adaptive Focused Acoustics) break method etc..Enzyme cutting method root
According to the selection of enzyme, two classes can be divided into again, one kind is to carry out DNA fragmentation using the restriction enzyme of specific restriction enzyme site,
Another kind of is to carry out random fracture to DNA by enzyme combination.Enzyme cutting method not only requires higher to sample purity, and to randomness
Limit.Adapted local cosine transform ultrasonic wave break method (AFA) is the method for ideal DNA fragmentation, especially Covaris
The instrument for the ultrasonic wave DNA breakage that company produces, there is the features such as fragmentation is accurate, and success rate is high.Commercially available another example, Billy
When Diagenode produce Biorupter, it is suitable with Covaris Products to DNA crushing effects.But above-mentioned two
Product price is at a relatively high, and experiment consumptive material is expensive, experimentation is complicated, experiment flux is relatively low, experimental facilities is safeguarded and changes generation
The factors such as valency height, it is set to be difficult to meet popular laboratory.
Therefore need a kind of new easily operated broken DNA method, can either precisely, high efficiency, high flux crush
Fragment, and can enough substantially reduce consumptive material, instrument cost.The utility model provides one kind and broken suitable for KQ-50E ultrasonic washing instruments
Broken DNA simple, practical device, can equivalent, convenient, fast, low input, carry out DNA to low consumption and crush.
Utility model content
The purpose of this utility model is in a kind of device that DNA is crushed suitable for high-flux sequence of offer.
The utility model includes following characteristics：
The device of broken DNA suitable for high-flux sequence a kind of, the device include square DNA sample fixed plate and surpassed
Sound wave cleaning device, offering in the plate face of the DNA sample fixed plate and put sample hole, described put is provided with heat emission hole around sample hole,
One jiao of the DNA sample fixed plate is set to positioning and smears angle, and groove on the rocks is further opened with described DNA sample fixed plate.
The DNA sample fixed plate top offers suppending hole.Its function is easy for suspension placement, ultrasonic disruption DNA
Shi Tianjia ice cubes.
The upper left corner that angle is located at the DNA sample fixed plate is smeared in described positioning.Described positioning smear angle for linear pattern or
Arc.Its function is to be used to prevent from placing mistake during the DNA sample fixed plate use.
Described sample hole of putting arranges for multiple and rectangular array or annular array.
Described put is provided with 6 heat emission holes around sample hole, being uniformly distributed in for described heat emission hole independence described puts sample hole
Around or be communicated in that described to put sample hole quincunx in 6 jiaos.The function of the heat emission hole is mainly used in reducing EP tube walls
Heat；The quantity of heat emission hole will excessively cause to put that sample is unstable, and crossing major general causes radiating effect bad.
The geomery of the DNA sample fixed plate matches with the ultrasonic response groove of the ultrasonic washing instrument.Institute
The material for stating DNA sample fixed plate is foam or plastics, but not limited to this.
Described groove on the rocks be located on the side of the DNA sample fixed plate or/and other three angles on.It is described on the rocks
The function of groove adds ice cube when being ultrasonic disruption DNA, to control water temperature.
In use, described DNA sample fixed plate is placed in described ultrasonic response groove, the DNA sample fixed plate
Put sample hole position in the surface zone of action of ultrasonic washing instrument transducer.
Described puts sample hole, and its position is located at the center of the device, is managed for placing the EP containing DNA to be broken；Its
Number and arrangement mode, can be 9 palace lattice, the hole of plum blossom 7, annular 6-8 holes etc.；
Application of the above-mentioned device in high-throughput sequencing library is built.
The application method of the above-mentioned broken DNA suitable for high-flux sequence device includes herein below with step：
(I) nucleic acid (DNA, RNA etc.) to be broken is added in EP pipes, and adjusts cumulative volume in EP pipes；
(II) putting step (I) the EP insertion DNA sample fixed plates in sample hole；
(III) H of certain volume is added into ultrasonic washing instrument2O, and adjust water temperature with trash ice；
(IV) the DNA sample fixed plate that EP pipes are inserted with step (II) Suo Shu is put into step (III) described ultrasonic washing instrument
In；
(V) setting time, open ultrasonic wave progress nucleic acid and crush；
The EP pipes that above-mentioned steps (I) use recommend 1.5mL EP pipes, if selecting the EP of other volumes to manage, it is ensured that selected EP pipes
Type matched with the sample hole of putting of the present apparatus；Population of samples product suggests 50-100uL, and volume is crossed the unmanageable ultrasonic wave of major general and broken
Broken effect, volume, which crosses conference, causes nucleic acid to crush heterogeneity；
When carrying out step (II) operation, it is ensured that EP manages the fully-inserted DNA sample fixed plate, checks the lid of all EP pipes
Whether sustained height is in；
Ultrasonic washing instrument described in step (III), it is recommended to use the KQ-50E single transducers of Russia and the U.S.'s electrical equipment, power 50W
Ultrasonic washing instrument；If use the ultrasonic washing instrument of similar brand, it is ensured that the parameter such as transducer, power is consistent；If use more
The ultrasonic washing instrument of individual 50W transducers, it is ensured that the sample hole position of putting of this DNA sample fixed plate is made in the surface of a certain transducer
With in region；And add the H of 8cm depth2O, regulation water temperature are 16 DEG C；
Time set by step (V), according to final nucleic acid want degree of crushing set, physical relationship approximately as：5-
DNA can be crushed to 500bp, 10min DNA can be crushed to 300bp, 20min and DNA can be crushed into 200bp by 7min；
In embodiment of the present utility model, the KQ-50E single transducers produced using Jiangsu company of Russia and the U.S., power 50W it is super
Sound wave cleaning device.
The broken DNA suitable for high-flux sequence of the utility model design device, there is advantages below and beneficial to effect
Fruit：
(1) it is easy that device is simple, consumptive material obtains；
(2) method is simple and feasible, is easy to actual execution；
(3) equivalent nucleic acid broken results can be obtained.
Brief description of the drawings
Fig. 1 is the structure chart of DNA sample fixed plate described in the utility model.
Fig. 2 is the structure chart of DNA sample fixed plate described in the utility model.
Fig. 3 is the structural representation of the device of the broken DNA described in the utility model suitable for high-flux sequence.In figure
The H of 8cm depth is added in the ultrasonic response groove of ultrasonic washing instrument2O, and water temperature is adjusted to 16 DEG C with trash ice, this practicality is new
DNA sample fixed plate designed by type is put on the water surface, with ultrasonic response groove perfect matching.
In Fig. 1~3,1-DNA sample fixed plates；Angle is smeared in 2- positioning；3- puts sample hole；4- heat emission holes；5- suppending holes；6- is on the rocks
Groove；7- ultrasonic response grooves, 8- transducers.
Fig. 4 is that the equipment that foreign Covaris companies provide carries out broken electrophoresis result figure to DNA, is shown respectively in figure
The effect of 180bp, 300bp, 500bp size.
Fig. 5 is to carry out DNA to 092 project sample using the device designed by the utility model to crush, and crushes time setting
For 7min electrophoresis result figure, show the segment ranges residing for DNA in 500bp in figure.
Fig. 6 is to carry out DNA to 062 project sample using the device designed by the utility model to crush, and crushes time setting
For 10min electrophoresis result figure, show the segment ranges residing for DNA in 300bp in figure.
Fig. 7 is to carry out DNA to 517 project samples using the device designed by the utility model to crush, and crushes time setting
For 20min (carried out during practical operation using 10min+10min modes, prevent single ultrasonication 20min produce amount of heat,
Influence experiment effect) electrophoresis result figure, show in figure the segment ranges residing for DNA in 200bp.
Embodiment
Following examples further illustrate the application effect of the device designed by the utility model, but should not be construed as to this
The limitation of utility model.In the case of without departing substantially from the utility model spirit and essence, to what is equipped mentioned by the utility model
The modifications or substitutions that each component is made, belong to the scope of the utility model.
Unless otherwise specified, the conventional meanses that technological means used in embodiment is well known to those skilled in the art；
Equipment used is commercial goods in embodiment.Ultrasonic equipment is KQ-50E single transducers, the work(that Jiangsu company of Russia and the U.S. produces
Rate 50W ultrasonic washing instrument.
Embodiment 1
As shown in Figure 1,2 and 3, a kind of broken DNA suitable for high-flux sequence device, the device include square
DNA sample fixed plate 1 and ultrasonic washing instrument, offer in the plate face of the DNA sample fixed plate 1 and put sample hole 3, it is described to put sample
Heat emission hole 4 is provided with around hole 3, the upper left corner of the DNA sample fixed plate 1 is set to linear positioning and smears angle 2 for preventing
The DNA sample fixed plate 1 places mistake when using, and groove 6 on the rocks is further opened with described DNA sample fixed plate 1.It is described
The top of DNA sample fixed plate 1 offer suppending hole 5 be used for hang placement or add ice cube during ultrasonic disruption DNA.
Described put is provided with 6 heat emission holes 4 around sample hole 3, independent being uniformly distributed in of described heat emission hole 4 described puts sample
Around hole (as shown in Figure 2) or to be communicated in described sample hole of putting be in 6 jiaos quincunx (as shown in Figure 1).The heat emission hole 4
Function is mainly used in reducing the heat of EP tube walls；The quantity of heat emission hole 4 will excessively cause to put that sample is unstable, and crossing major general causes to radiate
It is ineffective.
The geomery of the DNA sample fixed plate 1 matches with the ultrasonic response groove 7 of the ultrasonic washing instrument.
The material of the DNA sample fixed plate 1 is foam or plastics.
Described groove on the rocks 6 is located on the side and other three angles of the DNA sample fixed plate 1.It is located at the DNA
Groove on the rocks 6 on the side of sample fixed plate 1 is square, is located on the rocks recessed on other three angles of the DNA sample fixed plate 1
Groove 6 is circular arc.The function of the groove on the rocks 6 adds ice cube when being ultrasonic disruption DNA, to control water temperature.
In use, described DNA sample fixed plate 1 is placed in described ultrasonic response groove 7, the DNA sample is fixed
The sample hole 3 of putting of plate 1 is located in the surface zone of action of ultrasonic washing instrument transducer 8.Described its position of sample hole 3 of putting is located at
The center of the device, managed for placing the EP containing DNA to be broken；The quantity for putting sample hole 3 is 9 and rectangular array is arranged
Row.
The equipment provided by the utility model of embodiment 2 carries out broken application example to 092 project DNA
1. 092 project DNA 1ug to be broken are added in 1.5mL EP pipes, and use ddH2It is overall in O adjustment EP pipes
Product is 55uL；
2. 1.5mL EP pipes described in step 1 are inserted into the present apparatus to put in sample hole, and ensure to place reliable；
3. the H of certain volume is added into KQ-50E ultrasonic washing instruments2O, it is 8cm to make its depth of water, and is adjusted with trash ice
Water temperature；
4. the present apparatus that 1.5mL EP pipes are inserted with described in step 2 is put into ultrasonic washing instrument described in step 3；
5. setting time is 7min, opens ultrasonic wave progress nucleic acid and crush；
After 6. ultrasound terminates, 1.5mL EP pipes are taken out, it is of short duration that liquid in pipe is collected by centrifugation, take out 8uL progress agaroses and coagulate
Gel electrophoresis detect, and preserve picture (Fig. 5)；
7. by the result of comparison diagram 5 and Fig. 4, show that the present apparatus crushes DNA equivalent, side in high-flux sequence
Just, cheap advantage.
The equipment provided by the utility model of embodiment 3 carries out broken application example to 062 project DNA
1. 062 project DNA 1ug to be broken are added in 1.5mL EP pipes, and use ddH2It is overall in O adjustment EP pipes
Product is 55uL；
2. 1.5mL EP pipes described in step 1 are inserted into the present apparatus to put in sample hole, and ensure to place reliable；
3. the H of certain volume is added into KQ-50E ultrasonic washing instruments2O, it is 8cm to make its depth of water, and is adjusted with trash ice
Water temperature；
4. the present apparatus that 1.5mL EP pipes are inserted with described in step 2 is put into ultrasonic washing instrument described in step 3；
5. setting time is 10min, opens ultrasonic wave progress nucleic acid and crush；
After 6. ultrasound terminates, 1.5mL EP pipes are taken out, it is of short duration that liquid in pipe is collected by centrifugation, take out 8uL progress agaroses and coagulate
Gel electrophoresis detect, and preserve picture (Fig. 6)；
7. by the result of comparison diagram 6 and Fig. 4, illustrate that the present apparatus crushes DNA equivalent, side in high-flux sequence again
Just, cheap advantage.
The equipment provided by the utility model of embodiment 4 carries out broken application example to 517 project DNA
1. 517 projects DNA 1ug to be broken are added in 1.5mL EP pipes, and use ddH2It is overall in O adjustment EP pipes
Product is 55uL；
2. 1.5mL EP pipes described in step 1 are inserted into the present apparatus to put in sample hole, and ensure to place reliable；
3. the H of certain volume is added into KQ-50E ultrasonic washing instruments2O, it is 8cm to make its depth of water, and is adjusted with trash ice
Water temperature；
4. the present apparatus that 1.5mL EP pipes are inserted with described in step 2 is put into ultrasonic washing instrument described in step 3；
5. setting time is 10min, opens ultrasonic wave progress nucleic acid and crush；
After 6. ultrasound terminates, 1.5mL EP pipes are taken out, it is of short duration that liquid in pipe is collected by centrifugation, 1.5mL EP pipes are put into this dress
Put, repeat step 2-5 is once；
After 7. ultrasound terminates, 1.5mL EP pipes are taken out, it is of short duration that liquid in pipe is collected by centrifugation, take out 8uL progress agaroses and coagulate
Gel electrophoresis detect, and preserve picture (Fig. 7)；
8. by comparison diagram 7 and Fig. 4 result, further confirm the present apparatus crushed in high-flux sequence DNA it is equivalent,
Convenient, cheap advantage.
Although above having made detailed description to the utility model with a general description of the specific embodiments,
But on the basis of the utility model, it can be made some modifications or improvements, this is aobvious and easy to those skilled in the art
See.Therefore, the these modifications or improvements on the basis of without departing from the utility model spirit, belong to the utility model
Claimed scope.
Claims (10)
1. a kind of broken DNA suitable for high-flux sequence device, it is characterised in that the device includes square DNA sample
Fixed plate (1) and ultrasonic washing instrument, offer in the plate face of the DNA sample fixed plate and put sample hole (3), it is described to put sample hole
(3) it is provided with heat emission hole (4) around, one jiao of the DNA sample fixed plate (1) is set to positioning and smears angle (2), described DNA samples
Groove on the rocks (6) is further opened with product fixed plate (1).
2. device according to claim 1, it is characterised in that the top of the DNA sample fixed plate (1) offers suspension
Hole (5).
3. device according to claim 1, it is characterised in that described positioning, which smears angle (2) and is located at the DNA sample, to be fixed
The upper left corner of plate (1).
4. the device according to claim 1 or 3, it is characterised in that it is linear pattern or arc that angle (2) is smeared in described positioning.
5. device according to claim 1, it is characterised in that it is described put sample hole (3) for multiple and rectangular array or
Annular array arranges.
6. device according to claim 1 or 5, it is characterised in that described put is provided with 6 heat emission holes around sample hole (3)
(4) what, described heat emission hole (4) was independent be uniformly distributed in it is described put around sample hole (3) or be communicated in described put sample hole
(3) it is quincunx in 6 valves.
7. device according to claim 1, it is characterised in that the geomery of the DNA sample fixed plate (1) with it is described
The ultrasonic response groove (7) of ultrasonic washing instrument matches.
8. device according to claim 1, it is characterised in that the material of the DNA sample fixed plate (1) is foam or modeling
Material.
9. device according to claim 1, it is characterised in that described groove on the rocks (6) is located at the DNA sample and fixed
On the side of plate (1) or on other three angles.
10. device according to claim 1, it is characterised in that described groove on the rocks (6) is located at the DNA sample and consolidated
On on the side of fixed board (1) and other three angles.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720386522.9U CN206736220U (en) | 2017-04-13 | 2017-04-13 | A kind of device that DNA is crushed suitable for high-flux sequence |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720386522.9U CN206736220U (en) | 2017-04-13 | 2017-04-13 | A kind of device that DNA is crushed suitable for high-flux sequence |
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| Publication Number | Publication Date |
|---|---|
| CN206736220U true CN206736220U (en) | 2017-12-12 |
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ID=60560213
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|---|---|---|---|
| CN201720386522.9U Active CN206736220U (en) | 2017-04-13 | 2017-04-13 | A kind of device that DNA is crushed suitable for high-flux sequence |
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| Country | Link |
|---|---|
| CN (1) | CN206736220U (en) |
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2017
- 2017-04-13 CN CN201720386522.9U patent/CN206736220U/en active Active
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Denomination of utility model: Device suitable for broken DNA in high flux order -checking Effective date of registration: 20190926 Granted publication date: 20171212 Pledgee: Bank of China Limited by Share Ltd. Nanjing City South Branch Pledgor: JIANGSU GENESMILE PRECISION MEDICAL TECHNOLOGY CO.,LTD. Registration number: Y2019980000207 |
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Date of cancellation: 20200928 Granted publication date: 20171212 Pledgee: Bank of China Limited by Share Ltd. Nanjing City South Branch Pledgor: JIANGSU GENESMILE PRECISION MEDICAL TECHNOLOGY Co.,Ltd. Registration number: Y2019980000207 |
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Denomination of utility model: A device for DNA fragmentation in high throughput sequencing Effective date of registration: 20200929 Granted publication date: 20171212 Pledgee: Bank of China Limited by Share Ltd. Nanjing City South Branch Pledgor: JIANGSU GENESMILE PRECISION MEDICAL TECHNOLOGY Co.,Ltd. Registration number: Y2020980006613 |
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Date of cancellation: 20211012 Granted publication date: 20171212 Pledgee: Bank of China Limited by Share Ltd. Nanjing City South Branch Pledgor: JIANGSU GENESMILE PRECISION MEDICAL TECHNOLOGY Co.,Ltd. Registration number: Y2020980006613 |
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Denomination of utility model: A device suitable for breaking DNA in high-throughput sequencing Effective date of registration: 20211013 Granted publication date: 20171212 Pledgee: Bank of China Limited by Share Ltd. Nanjing City South Branch Pledgor: JIANGSU GENESMILE PRECISION MEDICAL TECHNOLOGY Co.,Ltd. Registration number: Y2021980010607 |
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