CN109609359A - Thallus liquid cracker and its cleavage method - Google Patents
Thallus liquid cracker and its cleavage method Download PDFInfo
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- CN109609359A CN109609359A CN201811592597.8A CN201811592597A CN109609359A CN 109609359 A CN109609359 A CN 109609359A CN 201811592597 A CN201811592597 A CN 201811592597A CN 109609359 A CN109609359 A CN 109609359A
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- Prior art keywords
- connecting tube
- thallus
- liquid
- way connection
- storage device
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/06—Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material
Abstract
The present invention designs field of biotechnology, specifically disclose a kind of thallus liquid cracker and its cleavage method, by thallus re-suspension liquid being uniformly mixed with lysate using pipeline instead of mixing manually, lytic effect reaches cracking phase same level manually, the difference and nonrepeatability generated when different operation person operation when avoiding manual mixing, simultaneously, on the basis of lytic effect is identical, whole cracking scale can be improved, manpower is saved by control conveying device conveying speed and the length of pipeline.
Description
Technical field
The present invention relates to field of biotechnology, and in particular to a kind of thallus liquid cracker and its cleavage method.
Background technique
Plasmid is the symbiotic type gene in bacterium, it can the vertical heredity and to assign host cell some in bacterium
Phenotype, plasmid are that foreign gene-carrying enters in bacterium the important medium for expanding or expressing, and the delivery vehicle of this gene is in base
Because having in engineering and wide application value.
Plasmid extraction mainly has alkaline lysis, boiling lysis and a small amount of one-step extractions etc., in the lab, uses alkali
Step usually requires to be manually added different lysate and neutralizer when cracking process extracts plasmid, during being manually added,
There is certain requirement to the dynamics and speed that are manually added, is the key point for influencing lytic effect in cracking operation, dynamics mistake
Big or overlong time can all lead to the reduction of plasmid supercoil ratio, and impurity value increases, meanwhile, in different cracking experiments, hand
Difference and nonrepeatability are easy to produce when different operation person operates when dynamic mixing.
Summary of the invention
For the above technical problems, the present invention provides a kind of thallus liquid cracker and its cleavage method.
In order to solve the above-mentioned technical problem, the technical solution adopted by the present invention are as follows:
A kind of thallus liquid cracker, including the first liquid storage device, the second liquid storage device, third liquid storage device, collection device, first
Three-way connection and the second three-way connection,
First liquid storage device and second liquid storage device pass through the first connecting tube and the second connecting tube and described the respectively
First and second interface of one three-way connection is connected, and is each provided with first in first connecting tube and second connecting tube
Conveying device and the second conveying device；
The third interface of first three-way connection and the third liquid storage device pass through third connecting tube and the 4th respectively and connect
Adapter tube is connected with the first and second interface of second three-way connection, and the third interface of second three-way connection also passes through the 5th
Connecting tube is connected with the collection device.
Further, first conveying device, the second conveying device and third conveying device are peristaltic pump.
Further, the length L of the third connecting tube is 1060mm-2120mm.
Based on above-mentioned thallus liquid cracker, the present invention also provides a kind of thallus liquid cleavage methods, including following step
It is rapid:
Step 1: pouring into thallus weight in first liquid storage device, second liquid storage device and the third liquid storage device respectively
Suspension, lysate and neutralizer；
Step 2: while starting first conveying device and the second conveying device, it is conveyed in first liquid storage device respectively
Thallus re-suspension liquid and second liquid storage device in lysate, enter first each by the first connecting tube and the second connecting tube
Three-way connection, and tentatively mixed inside first three-way connection, obtain mixed liquor；
Step 3: starting the third conveying device, convey the neutralizer in the third liquid storage device, pass through the 4th connection
The second interface that pipe reaches the second three-way connection goes out, and closes third conveying device；
Step 4: the mixed liquor in step 2 is entered by the third connecting tube reaches the first of second three-way connection
When interface, start the third conveying device, so that mixed liquor and neutralizer is entered the second three-way connection, and the described 2nd 3
It is mixed again inside pass joint, obtains secondary mixed liquor；
Step 5: secondary mixed liquor is entered in the collection device by the 5th connecting tube.
Further, first connecting tube is identical as the length of the second connecting tube；First conveying device and second defeated
Send the conveying speed of device identical.
The beneficial effects of the present invention are: the present invention provides a kind of thallus liquid cracker and its cleavage methods, pass through
Thallus liquid cracker, which mixes thallus re-suspension liquid with lysate, replaces hand mix, and mixing is more uniform, and lytic effect reaches
Cracking phase same level manually, the difference and nonrepeatability generated when different operation person operates when avoiding hand mix, meanwhile,
On the basis of lytic effect is identical, by control conveying device conveying speed and the length of connecting tube, the thallus liquid is promoted
The efficiency of cracker, it is high-efficient, stability is good to improve whole cracking reaction scale, save manpower.
Detailed description of the invention
It, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical solution in the prior art
Embodiment or attached drawing needed to be used in the description of the prior art are briefly described.In all the appended drawings, similar element
Or part is generally identified by similar appended drawing reference.In attached drawing, each element or part might not be drawn according to actual ratio.
Fig. 1 is the structural schematic diagram of thallus liquid cracker of the present invention.
Specific embodiment
With reference to the accompanying drawing, the present invention is described in detail.
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to the accompanying drawings and embodiments, right
The present invention is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, and
It is not used in the restriction present invention.
Embodiment 1
It is an example structure schematic diagram of thallus liquid cracker of the invention, arrow indicates liquid in Fig. 1 referring to Fig. 1
Body flow direction, specifically, the thallus liquid cracker of the present embodiment, including the first liquid storage device 11, the second liquid storage device 12, third
Liquid storage device 13, collection device 2, the first three-way connection 31 and the second three-way connection 32, wherein
First liquid storage device 11 and the second liquid storage device 12 pass through the first connecting tube 41 and the second connecting tube 42 and the one or three respectively
First and second interface of pass joint 31 is connected, and the first conveying dress is each provided in the first connecting tube 41 and the second connecting tube 42
Set 51 and second conveying device 52；
The third interface and third liquid storage device 13 of first three-way connection 31 pass through third connecting tube 43 and the 4th connection respectively
Pipe 44 is connected with the first and second interface of the second three-way connection 32, and the third interface of the second three-way connection 32 also passes through the 5th connection
Pipe 45 is connected with collection device 2.
In the present embodiment, thallus weight is poured into respectively in the first liquid storage device 11, the second liquid storage device 12 and third liquid storage device 13
Suspension, lysate and neutralizer restart the first conveying device 51, convey the thallus re-suspension liquid in the first liquid storage device 11, so that
Thallus re-suspension liquid is entered in the first three-way connection 31 by the first connecting tube 41 from the first interface of the first three-way connection 31；Together
When, start the second conveying device 52, convey the second liquid storage device 12 in lysate so that lysate by the second connecting tube 42 from
The second interface of first three-way connection 31 enters in the first three-way connection 31；And thallus re-suspension liquid and lysate reach the one or three
It can tentatively mix and react behind 31 inside of pass joint, then, mixed liquor enters the from the third interface of the first three-way connection 31
In three connecting tubes 43, at this point, moving since mixed liquor relays afterflow in third connecting tube 43, thallus re-suspension liquid and lysate are increased
Contact area, so that thallus re-suspension liquid can be made uniformly to mix, react with lysate；When mixed liquor is from the first three-way connection 31
Third interface when entering in third connecting tube 43, while starting third conveying device 53, convey in third liquid storage device 13
And liquid, neutralizer are reached by the 4th connecting tube 44 at the second interface of second three-way connection 32, and third conveying device 53 is closed；
After mixed liquor enters in third connecting tube 43, since the length of third connecting tube 43 is longer, when mixed in third connecting tube 43
When closing at the first interface of liquid the second three-way connection 32 of arrival, it is again started up third conveying device 53, thus the second three-way connection
Neutralizer at 32 second interface enters the two or three with the mixed liquor at the first interface of the second three-way connection 32 simultaneously and connects
It inside first 32, mixes mixed liquor with neutralizer and reacts, obtain secondary mixed liquor；Then, secondary mixed liquor is from the two or three
The third interface of pass joint 32 enter the 5th connecting tube 45 in, continue in the 5th connecting tube 45 in and；Finally, secondary mixed
Liquid is closed to enter in collection device 2.
Further, in this embodiment the first conveying device 51, the second conveying device 52 and third conveying device 53 are
Peristaltic pump.In the present embodiment, pass through the pump speed of the first conveying device 51 of control, the second conveying device 52 and third conveying device 53
The effect of reaction is controlled, the difference and nonrepeatability generated when different operation person operates when avoiding hand mix, thus
Whole cracking operation scale is improved, the efficiency of the thallus liquid cracker is improved, and saves manpower.
Further, in the present embodiment, the length L of third connecting tube 43 is 1060mm-2120mm.
In the present embodiment, the flow velocity for setting the first connecting tube 41 and the second connecting tube 42 is 1.0ml/s, thallus re-suspension liquid
It is 2min, 3min, 4min with the pyrolysis time of lysate, the first connecting tube 41, the second connecting tube 42, third connecting tube 43, the
Four connecting tubes 44 and the 5th connecting tube 45 are all made of the internal diameter of same specification, and measurement obtains internal diameter r=0.6cm,
Pass through formula: (π r2× L)/1.0ml/s=tTime
Pyrolysis time is brought into formula to be calculated: the length L of third connecting tube 43 be respectively 1060mm, 1590mm and
2120mm。
Embodiment 2
Based on above-mentioned thallus liquid cracker, the present invention also provides a kind of thallus liquid cleavage methods, including following step
It is rapid:
Step 1: poured into the first liquid storage device 11, the second liquid storage device 12 and third liquid storage device 13 respectively thallus re-suspension liquid,
Lysate and neutralizer；
Step 2: while starting the first conveying device 51 and the second conveying device 52, it is conveyed in the first liquid storage device 11 respectively
Lysate in thallus re-suspension liquid and the second liquid storage device 12, so that thallus re-suspension liquid and lysate are each by the first connecting tube 41
Enter the first three-way connection 31 with the second connecting tube 42, and tentatively mixed inside the first three-way connection 31, is mixed
Liquid；
Step 3: starting third conveying device 53 conveys the neutralizer in third liquid storage device 13, passes through the 4th connecting tube 44
The second interface for reaching the second three-way connection 32 goes out, and closes third conveying device 53；
Step 4: the mixed liquor in step 2 enters the first interface for reaching the second three-way connection 32 by third connecting tube 43
When place, start third conveying device 53, so that mixed liquor and neutralizer is entered the second three-way connection 32, and in the second three-way connection 32
Inside is mixed again, obtains secondary mixed liquor；
Step 5: secondary mixed liquor is entered in collection device 2 by the 5th connecting tube 45.
In the present embodiment, the first connecting tube 41 is identical as the length of the second connecting tube 42；First conveying device 51 and second
The conveying speed of conveying device 52 is identical.
In the present embodiment, since the first connecting tube 41 and the second connecting tube 42 connect with the first of the first three-way connection 31 respectively
Mouth, second interface connection, if the conveying speed of the thallus re-suspension liquid in the first connecting tube 41 is than the cracking in the second connecting tube 42
The conveying speed of liquid is fast, then a part of thallus re-suspension liquid can first pass through the first three-way connection 31, and can be from the first three-way connection 31
Third interface enter in third connecting tube 43, at this point, cannot carry out reacting with lysate will for this part thallus re-suspension liquid
It enters in the second three-way connection 32, due to this part, thallus re-suspension liquid is not mixed with lysate, reacted, then is entirely split
Solution preocess will be incomplete, so that lytic effect be not achieved；If the conveying speed ratio of the thallus re-suspension liquid in the first connecting tube 41
The conveying speed of lysate in second connecting tube 42 is slow, then a part of lysate can first pass through the first three-way connection 31, and meeting
Enter in third connecting tube 43 from the third interface of the first three-way connection 31, fraction of lysate can remain in third connecting tube 43
In, at this point, the thallus re-suspension liquid in the first connecting tube 41 is passed through with the lysate in the second connecting tube 42 in the first three-way connection 31
After crossing preliminary mixing, enters in third connecting tube 43 from the third interface of the first three-way connection 31, will continue to and remain in third
Lysate in connecting tube 43 is mixed, is reacted, also, due to the conveying speed of the lysate in the second connecting tube 42 compared with
Fastly, lysate enter the amount in third connecting tube 43 from the third interface of the first three-way connection 31 will be than thallus re-suspension liquid more
It is more, thus cracking reaction can be made excessive, so that the lytic effect of needs be not achieved.
Therefore, in order to guarantee cracking reaction sufficiently, uniformly, the first connecting tube 41, second connection used in the present embodiment
Pipe 42, third connecting tube 43, the 4th connecting tube 44 are identical with the bore of the 5th connecting tube 45, and the first connecting tube 41 and second connects
The length of adapter tube 42 is identical, and is in the same horizontal position；The conveying of first conveying device 51 and the second conveying device 52 is set
Speed is identical, and at the same time starting, to guarantee the thallus re-suspension liquid and the second connecting tube in the first connecting tube 41 of accurate control
The conveying speed of lysate in 42 is identical, and then ensures the thallus re-suspension liquid in the first connecting tube 41 and the second connecting tube 42
In lysate can reach simultaneously in the first three-way connection 31, make thallus re-suspension liquid mixed with lysate sufficiently, guarantee cracking effect
Fruit.
Hand mix is replaced by mixing thallus re-suspension liquid with lysate using thallus liquid cracker, mixing, reaction
More uniform, lytic effect reaches cracks identical level manually, generation when different operation person operates when avoiding hand mix
Difference and nonrepeatability, meanwhile, on the basis of lytic effect is identical, the conveying of each conveying device of control can be passed through
The length of speed and connecting tube, improves whole cracking scale, high-efficient, stability is good, saves manpower.
The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；Although referring to aforementioned each reality
Applying example, invention is explained in detail, those skilled in the art should understand that: it still can be to aforementioned each
Technical solution documented by embodiment is modified, or equivalent substitution of some or all of the technical features；And
These are modified or replaceed, the range for technical solution of various embodiments of the present invention that it does not separate the essence of the corresponding technical solution,
It should all cover within the scope of the claims and the description of the invention.
Claims (5)
1. a kind of thallus liquid cracker, it is characterised in that: including the first liquid storage device, the second liquid storage device, third liquid storage device, collect
Device, the first three-way connection and the second three-way connection,
First liquid storage device and second liquid storage device pass through the first connecting tube and the second connecting tube and the described 1st respectively
First and second interface of pass joint is connected, and is each provided with the first conveying in first connecting tube and second connecting tube
Device and the second conveying device；
The third interface of first three-way connection and the third liquid storage device pass through third connecting tube and the 4th connecting tube respectively
It is connected with the first and second interface of second three-way connection, the third interface of second three-way connection also passes through the 5th connection
Pipe is connected with the collection device.
2. thallus liquid cracker according to claim 1, it is characterised in that: first conveying device, the second conveying
Device and third conveying device are peristaltic pump.
3. thallus liquid cracker according to claim 1, it is characterised in that: the length L of the third connecting tube is
1060mm-2120mm。
4. a kind of thallus liquid cleavage method uses thallus liquid cracker described in claim 1-3 any one claim
It is cracked, characterized in that it comprises the following steps:
Step 1: pouring into thallus resuspension in first liquid storage device, second liquid storage device and the third liquid storage device respectively
Liquid, lysate and neutralizer；
Step 2: while starting first conveying device and the second conveying device, the bacterium in first liquid storage device is conveyed respectively
Lysate in weight suspension and second liquid storage device, so that the thallus re-suspension liquid and the lysate are each by first
Connecting tube and the second connecting tube enter the first three-way connection, and are tentatively mixed inside first three-way connection, obtain
Mixed liquor；
Step 3: starting the third conveying device, convey the neutralizer in the third liquid storage device, arrived by the 4th connecting tube
Up at the second interface of the second three-way connection, third conveying device is closed；
Step 4: the mixed liquor in step 2 enters the first interface for reaching second three-way connection by the third connecting tube
When place, it is again started up the third conveying device, so that mixed liquor and neutralizer is entered the second three-way connection, and the described 2nd 3
It is mixed again inside pass joint, obtains secondary mixed liquor；
Step 5: secondary mixed liquor is entered in the collection device by the 5th connecting tube.
5. thallus liquid cleavage method according to claim 4, it is characterised in that: first connecting tube and the second connecting tube
Length it is identical；First conveying device is identical as the conveying speed of the second conveying device.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201811592597.8A CN109609359A (en) | 2018-12-25 | 2018-12-25 | Thallus liquid cracker and its cleavage method |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811592597.8A CN109609359A (en) | 2018-12-25 | 2018-12-25 | Thallus liquid cracker and its cleavage method |
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| CN201811592597.8A Pending CN109609359A (en) | 2018-12-25 | 2018-12-25 | Thallus liquid cracker and its cleavage method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111979109A (en) * | 2020-09-01 | 2020-11-24 | 深圳普瑞金生物药业有限公司 | Plasmid vector continuous cracking device |
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| CN205687904U (en) * | 2016-06-17 | 2016-11-16 | 博奥生物集团有限公司 | A kind of micro fluidic device for nucleic acid extraction and control system |
| CN207933420U (en) * | 2016-07-06 | 2018-10-02 | 广州好芝生物科技有限公司 | A kind of micro-fluidic chip of genetic test |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6664049B1 (en) * | 1999-01-20 | 2003-12-16 | Aventis Pasteur S.A. | Method and device for cell lysis |
| CN1710057A (en) * | 2005-06-22 | 2005-12-21 | 殷玉和 | Multi-stage cell rupture and plasmid crude extracting reactor |
| CN1948481A (en) * | 2006-10-26 | 2007-04-18 | 中国农业大学 | Method of continuous large quantity extracting plasmid |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111979109A (en) * | 2020-09-01 | 2020-11-24 | 深圳普瑞金生物药业有限公司 | Plasmid vector continuous cracking device |
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