CN209854124U - Whole-sea deep microorganism nucleic acid in-situ extraction device - Google Patents
Whole-sea deep microorganism nucleic acid in-situ extraction device Download PDFInfo
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- CN209854124U CN209854124U CN201920431957.XU CN201920431957U CN209854124U CN 209854124 U CN209854124 U CN 209854124U CN 201920431957 U CN201920431957 U CN 201920431957U CN 209854124 U CN209854124 U CN 209854124U
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- 244000005700 microbiome Species 0.000 title claims abstract description 40
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 37
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 37
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 37
- 238000000605 extraction Methods 0.000 title claims abstract description 31
- 238000011065 in-situ storage Methods 0.000 title claims abstract description 28
- 239000013535 sea water Substances 0.000 claims abstract description 60
- 239000012528 membrane Substances 0.000 claims abstract description 37
- 230000002572 peristaltic effect Effects 0.000 claims abstract description 22
- 238000001179 sorption measurement Methods 0.000 claims abstract description 16
- 239000006166 lysate Substances 0.000 claims abstract description 15
- 239000012530 fluid Substances 0.000 claims description 8
- 238000001914 filtration Methods 0.000 abstract description 10
- 230000014509 gene expression Effects 0.000 abstract description 7
- 230000007613 environmental effect Effects 0.000 abstract description 3
- 230000000813 microbial effect Effects 0.000 abstract description 3
- 230000006037 cell lysis Effects 0.000 abstract description 2
- 239000002253 acid Substances 0.000 abstract 1
- 230000003834 intracellular effect Effects 0.000 abstract 1
- 108090000623 proteins and genes Proteins 0.000 description 11
- 239000000523 sample Substances 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 5
- 238000000034 method Methods 0.000 description 4
- 230000009089 cytolysis Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000005070 sampling Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 230000002934 lysing effect Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 238000012827 research and development Methods 0.000 description 2
- 241001131796 Botaurus stellaris Species 0.000 description 1
- 108091005461 Nucleic proteins Proteins 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000012165 high-throughput sequencing Methods 0.000 description 1
- 230000002706 hydrostatic effect Effects 0.000 description 1
- 210000004020 intracellular membrane Anatomy 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000009931 pascalization Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000004952 protein activity Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The utility model provides a whole-sea deep microorganism nucleic acid in-situ extraction device, after the in-situ extraction device along with deep sea platform dive to the deep sea, through the operation of sea water pump, pour into a large amount of sea water into the filter membrane chamber and accomplish microorganism enrichment filtration, microorganism in the sea water is filtered on the filter membrane, accomplish after filtering operation, open deep sea peristaltic pump, the lysate is poured into the filter membrane chamber, after a period of stillness, the microbial cell lysis on the filter membrane, the intracellular core acid dissolves in the lysate, open tee bend electromagnetic directional valve, inject the lysate that contains nucleic acid on the adsorption column, accomplish microorganism and larva nucleic acid DNA and RNA adsorption extraction in the deep sea in situ, compared with the prior art, the nucleic acid information of deep sea in-situ extraction avoids the influence of environmental changes such as temperature, pressure to nucleic acid information, the pollution of laboratory extraction process has also been avoided, the state under the sample in-situ environment has been reduced, the gene expression information of deep-sea microorganisms is truly kept, and valuable samples are provided for subsequent omics analysis.
Description
Technical Field
The utility model relates to a deep sea microorganism draws technical field, in particular to full sea deep microorganism nucleic acid normal position extraction element.
Background
Deep sea organisms play an important role throughout the ocean and even in the global ecosystem. Deep sea environments are typically characterized by high hydrostatic pressure and low temperature, and there are extreme environments such as deep sea hot water, cold springs, and bittern pools. Due to the ecological stress, deep-sea organisms must produce genes, proteins and secondary metabolites with special functions in the process of adapting to the extreme environment of deep sea. In the past years, the genome data of deep sea organisms are increased explosively due to the application of high-throughput sequencing technology, but the function of the genes of the deep sea organisms is still far behind, and huge gene resources contained in the data are still to be developed. The functional gene refers to a DNA fragment with definite physiological and biochemical activity and expression regulation and control mechanism. Due to insufficient support of effective deep sea detection equipment, serious obstacles exist in the research of deep sea biological functional genes.
In conventional sampling, after the sample is collected, subsequent processing is performed on the vessel. In the process, the deep sea organisms are influenced by factors such as temperature, salinity and hydrostatic pressure, and have great influence on the specific gene expression products in the deep sea. Some organisms can not restore the expression information and protein activity characteristics of functional genes in the in-situ environment of a sample due to cell rupture caused by sudden pressure reduction. Although some sampling equipment for deep-sea organisms have been developed at home and abroad, particularly, the research and development of the microorganism in-situ pressure-maintaining sampling device opens up a new place for the research on functional genes of the whole deep-sea organisms, the research on the specific functional genes of the deep sea cannot be deeply carried out due to the factors of small capacity, no heat preservation and the like of the pressure-maintaining sampler. In order to accurately detect the expression and action activity of deep-sea organism functional genes and discover a large number of functional genes of deep-sea organisms, the research and development of in-situ biological sample treatment and detection technology are imperative.
SUMMERY OF THE UTILITY MODEL
Therefore, there is a need to provide an in-situ extraction device for nucleic acid from deep microorganisms in whole sea, which can extract, collect and store the nucleic acid in situ.
In order to achieve the above purpose, the utility model adopts the following technical scheme:
an in-situ extraction device for nucleic acid of full-sea deep microorganisms comprises: sea water pump, peristaltic pump, check valve, first three-way valve, flowmeter, second three-way valve, filter membrane chamber, tee bend solenoid directional valve and adsorption column, sea water pump the check valve the first three-way valve the flowmeter the second three-way valve the filter membrane chamber the tee bend solenoid directional valve reaches the adsorption column passes through the pipeline and connects gradually, the peristaltic pump is connected the entry of check valve, wherein:
when the in-situ extraction device is submerged to the deep sea, the seawater pump injects seawater, the seawater enters the filter membrane cavity through the one-way valve, the first three-way valve, the flowmeter and the second three-way valve, the filter membrane cavity filters the flowing seawater, and microorganisms and larvae in the seawater are filtered on the filter membrane of the filter membrane cavity;
the flow meter detects the flow of the filtered seawater and obtains the total amount of the filtered seawater, when the filtered seawater meets the total amount requirement, the seawater pump is closed and the peristaltic pump is started, and the pre-prepared lysate is injected into the filter membrane cavity through the one-way valve, the first three-way valve, the flow meter and the second three-way valve;
and dissociating nucleic acid generated after the microorganisms and the larva cells on the filter membrane are lysed in the lysate, opening the three-way electromagnetic directional valve and the peristaltic pump, and injecting the lysate containing the nucleic acid into the adsorption column.
In some preferred embodiments, the external controller can adjust the power of the seawater pump and the peristaltic pump according to the seawater flow measured by the flow meter.
In some preferred embodiments, the sea water pump further comprises a pressure sensor connected to the third path of the second three-way valve, the pressure sensor is used for detecting the fluid pressure of the liquid pipeline, and the external controller is used for controlling and adjusting the driving force of the sea water pump and the peristaltic pump through the fluid pressure.
The utility model adopts the above technical scheme's advantage is:
the utility model provides a pair of whole sea deep microorganisms nucleic acid normal position extraction element, include: the method comprises the following steps of injecting a large amount of seawater into a filter membrane cavity to finish microorganism enrichment and filtration by the operation of a seawater pump after an in-situ extraction device is submerged to the deep sea along with a deep sea platform, opening the deep sea peristaltic pump after the filtering operation is finished, injecting lysis solution into the filter membrane cavity, after standing for a period of time, lysing microorganism cells on the filter membrane, dissolving contents such as nucleic acid, protein and the like in the cells into the lysis solution, opening a three-way electromagnetic directional valve, injecting the lysis solution containing nucleic acid onto an adsorption column, completing the adsorption and extraction of DNA and RNA of the microorganism and larva in situ, and compared with the prior art, the nucleic acid information extracted in situ avoids the influence of environmental changes such as temperature, pressure and the like on the nucleic acid information, the pollution of the extraction process of a laboratory is avoided, the state of the sample in the in-situ environment is restored, the gene expression information of the deep-sea microorganisms is kept most truly, and a valuable sample is provided for the subsequent omics analysis.
Furthermore, the utility model provides a full sea depth microorganism nucleic acid normal position extraction element, normal position microorganism enrichment, filtration compare traditional sea water sample mode, can short time, once only acquire more sea water to obtain more microorganism samples.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic structural diagram of an in-situ extraction apparatus for nucleic acid from microorganisms in whole sea depth according to an embodiment of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative efforts belong to the protection scope of the present invention.
Referring to fig. 1, the present invention provides an in-situ extraction apparatus for nucleic acid from deep-sea microorganisms, comprising: the device comprises a seawater pump 1, a peristaltic pump 2, a one-way valve 3, a first three-way valve 4, a flowmeter 5, a second three-way valve 7, a filter membrane cavity 8, a three-way electromagnetic directional valve 9 and an adsorption column 10. Wherein:
sea water pump 1, check valve 3 first three-way valve 4 the flowmeter 5 the second three-way valve 7 the filter membrane chamber 8 the tee bend solenoid directional valve 9 reaches adsorption column 10 connects gradually through the pipeline, peristaltic pump 2 is connected the entry of check valve 3.
In some preferred embodiments, the seawater pump 1, the peristaltic pump 2, the one-way valve 3, the first three-way valve 4, the flow meter 5, the second three-way valve 7, the filter membrane cavity 8, the three-way electromagnetic directional valve 9 and the adsorption column 10 all adopt an oil compensation form, and are packaged independently, so that the seawater pump can be exposed to deep sea water to work, the in-situ extraction process of microbial nucleic acid is ensured, and pipelines have no compensation oil leakage pollution.
In some preferred embodiments, the filter membrane chamber 8 is modular in design, easy to assemble and disassemble, and can be used for enriching and lysing organisms with different sizes by using mesh filter membranes with different sizes.
The utility model provides a deep microorganism nucleic acid normal position extraction element in full sea, working method is as follows:
when the whole-sea deep-microorganism nucleic acid in-situ extraction device submerges to the deep sea along with a deep sea platform (lander, ROV and the like), an external controller sends a command to control a seawater pump 1 to work, the seawater pump 1 injects seawater, the seawater enters a filter membrane cavity 8 through a one-way valve 3, a first three-way valve 4, a flowmeter 5 and a second three-way valve 7, the filter membrane cavity 8 filters the seawater flowing through, and microorganisms and larvae in the seawater are filtered on a filter membrane of the filter membrane cavity 8;
the flow meter 5 detects the flow of the filtered seawater and obtains the total amount of the filtered seawater, when the filtered seawater meets the total amount requirement, the seawater pump 1 is closed and the peristaltic pump 2 is opened, and the pre-configured lysate is injected into the filter membrane cavity 8 through the one-way valve 3, the first three-way valve 7, the flow meter 5 and the second three-way valve 7. It can be understood that the flow meter 5 accurately controls the flow of the fluid, the flow meter 5 measures the flow of the seawater, the external controller in signal connection with the flow meter 5 obtains flow feedback, the power of the seawater pump 1 and the peristaltic pump 2 is automatically adjusted, and the flow is accurately controlled; meanwhile, the flowmeter 5 can obtain the total flow, and when the seawater filtering reaches the preset total flow, the external controller can automatically control to complete the filtering operation.
Nucleic acid generated after the microbes and the larva cells on the filter membrane are cracked is dissociated in the lysate, the three-way electromagnetic directional valve 9 and the peristaltic pump 2 are opened, and the lysate containing the nucleic acid is injected into the adsorption column 10.
In some preferred embodiments, the system further comprises a pressure sensor 6, the pressure sensor 6 is connected to the third path of the second three-way valve 7, the pressure sensor 6 is used for detecting the fluid pressure of the liquid line, and an external controller adjusts the driving force of the seawater pump 1 and the peristaltic pump 2 through the fluid pressure control to control the line pressure within a reasonable range.
The utility model provides a deep microorganism nucleic acid normal position extraction element in whole sea, after normal position extraction element dives to the deep sea along with the deep sea platform, through the operation of sea water pump, pour into a large amount of sea water into the filter membrane chamber and accomplish microorganism enrichment filtration, microorganism in the sea water is filtered on the filter membrane, accomplish after filtering operation, open deep sea peristaltic pump, the lysate is poured into the filter membrane chamber, after a period of stillness, the microbial cell lysis on the filter membrane, intracellular nucleic acid, contents such as albumen dissolve in the lysate, open tee bend electromagnetic directional valve, inject the lysate that contains nucleic acid onto the adsorption column, accomplish microorganism and larva nucleic acid DNA and RNA adsorption extraction in deep sea normal position, compared with the prior art, the nucleic acid information that the deep sea normal position was drawed avoids the influence of environmental changes such as temperature, pressure to nucleic acid information, the pollution of laboratory extraction process has also been avoided, the state of the sample in the in-situ environment is restored, the gene expression information of the deep-sea microorganisms is truly reserved, and a valuable sample is provided for the subsequent omics analysis.
Furthermore, the utility model provides a full sea depth microorganism nucleic acid normal position extraction element, normal position microorganism enrichment, filtration compare traditional sea water sample mode, can short time, once only acquire more sea water to obtain more microorganism samples.
Of course, the whole-sea-depth microorganism nucleic acid in-situ extraction device of the present invention may have various changes and modifications, and is not limited to the specific structure of the above embodiment. In conclusion, the scope of the present invention should include those changes or substitutions and modifications which are obvious to those of ordinary skill in the art.
Claims (3)
1. An in-situ extraction device for nucleic acid of full-sea-depth microorganisms is characterized by comprising: sea water pump, peristaltic pump, check valve, first three-way valve, flowmeter, second three-way valve, filter membrane chamber, tee bend solenoid directional valve and adsorption column, sea water pump the check valve the first three-way valve the flowmeter the second three-way valve the filter membrane chamber the tee bend solenoid directional valve reaches the adsorption column passes through the pipeline and connects gradually, the peristaltic pump is connected the entry of check valve, wherein:
when the in-situ extraction device is submerged to the deep sea, the seawater pump injects seawater, the seawater enters the filter membrane cavity through the one-way valve, the first three-way valve, the flowmeter and the second three-way valve, the filter membrane cavity filters the flowing seawater, and microorganisms and larvae in the seawater are filtered on the filter membrane of the filter membrane cavity;
the flow meter detects the flow of the filtered seawater and obtains the total amount of the filtered seawater, when the filtered seawater meets the total amount requirement, the seawater pump is closed and the peristaltic pump is started, and the pre-prepared lysate is injected into the filter membrane cavity through the one-way valve, the first three-way valve, the flow meter and the second three-way valve;
and dissociating nucleic acid generated after the microorganisms and the larva cells on the filter membrane are lysed in the lysate, opening the three-way electromagnetic directional valve and the peristaltic pump, and injecting the lysate containing the nucleic acid into the adsorption column.
2. The in situ extraction apparatus for nucleic acid from deep microorganisms in whole sea according to claim 1, wherein an external controller is capable of adjusting the power of the seawater pump and the peristaltic pump according to the flow rate of seawater measured by the flow meter.
3. The in situ extraction apparatus for nucleic acid from deep sea microorganisms according to claim 1, further comprising a pressure sensor connected to the third path of the second three-way valve, wherein the pressure sensor is used for detecting the fluid pressure in the fluid line, and the external controller controls and adjusts the driving force of the sea water pump and the peristaltic pump according to the fluid pressure.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201920431957.XU CN209854124U (en) | 2019-04-01 | 2019-04-01 | Whole-sea deep microorganism nucleic acid in-situ extraction device |
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| CN201920431957.XU CN209854124U (en) | 2019-04-01 | 2019-04-01 | Whole-sea deep microorganism nucleic acid in-situ extraction device |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114540185A (en) * | 2022-03-10 | 2022-05-27 | 浙江大学 | Marine microorganism high-flux in-situ filtering device based on multi-channel circulating distributor |
| CN117568143A (en) * | 2024-01-15 | 2024-02-20 | 山东省海洋科学研究院(青岛国家海洋科学研究中心) | Deep sea sample nucleic acid in-situ digestion and preservation device |
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2019
- 2019-04-01 CN CN201920431957.XU patent/CN209854124U/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114540185A (en) * | 2022-03-10 | 2022-05-27 | 浙江大学 | Marine microorganism high-flux in-situ filtering device based on multi-channel circulating distributor |
| CN117568143A (en) * | 2024-01-15 | 2024-02-20 | 山东省海洋科学研究院(青岛国家海洋科学研究中心) | Deep sea sample nucleic acid in-situ digestion and preservation device |
| CN117568143B (en) * | 2024-01-15 | 2024-03-22 | 山东省海洋科学研究院(青岛国家海洋科学研究中心) | Deep sea sample nucleic acid in-situ digestion and preservation device |
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