CN103484354B - Nucleic acid extraction chip capable of extracting nucleic acid of gram-positive bacteria and gram-negative bacteria - Google Patents
Nucleic acid extraction chip capable of extracting nucleic acid of gram-positive bacteria and gram-negative bacteria Download PDFInfo
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Abstract
The invention discloses a nucleic acid extraction chip capable of extracting nucleic acid of gram-positive bacteria and gram-negative bacteria. The nucleic acid extraction chip comprises a control channel layer, an elastic thin film layer and a liquid channel layer; the upper surface of the liquid channel layer is provided with a nucleic acid extracting solution inlet channel (6), a rinsing solution inlet channel (5), an EDTA SDS inlet channel (4), a nuclease inlet channel (3), a cell lysis solution inlet channel (2) and a cell suspension inlet channel (1); the left ends of the nucleic acid extracting solution inlet channel (6), the rinsing solution inlet channel (5), the EDTA SDS inlet channel (4), the nuclease inlet channel (3), the cell lysis solution inlet channel (2) and the cell suspension inlet channel (1) are respectively provided with a liquid inlet which penetrates the liquid channel layer downward. The nucleic acid extraction chip capable of extracting nucleic acid of gram-positive bacteria and gram-negative bacteria is based on silicon bead test, centrifugation treatment is not needed, the nucleic acid extraction chip is convenient for automation operation, operation is simple and rapid, detection is fast, and detection effect is objective.
Description
Technical field
The present invention relates to a kind of nucleic acid extraction chip extracting gram-positive bacteria and Gram-negative bacteria nucleic acid.
Background technology
The extraction of nucleic acid relates to a basic fundamental of biological operation.Up to now, people have developed multiple method to extract nucleic acid.But wherein great majority relate to centrifugal, this is unfavorable for the microminiaturization and the automation that realize nucleic acid extraction.
Summary of the invention
One is the object of the present invention is to provide not need through centrifugal treating, be convenient to automation mechanized operation, simple, convenient, detect rapidly, effect is objective, can be the nucleic acid extraction chip extracting gram-positive bacteria and Gram-negative bacteria nucleic acid based on silica bead method that scientific research, medical diagnosis on disease, genetic test, forensic identification etc. provide technical support.
The present invention extracts the nucleic acid extraction chip of gram-positive bacteria and Gram-negative bacteria nucleic acid, comprises control channel layer, elastic film layer and fluid passage layer, the lower surface of control channel layer and the upper surface of elastic film layer are close to setting, the lower surface of elastic film layer and the upper surface of described fluid passage layer are close to setting, and the left side of described fluid passage layer upper surface is provided with in left-right direction from front to back successively into nucleic acid extraction liquid passage, enter rinsing liquid passage, enter EDTA SDS passage, enter nuclease passage, enter cell pyrolysis liquid passage and enter cell suspension passage, entering nucleic acid extraction liquid passage, enter rinsing liquid passage, enter EDTA SDS passage, enter nuclease passage, enter cell pyrolysis liquid passage and be respectively equipped with the left end entering cell suspension passage the inlet penetrating described fluid passage layer downwards, enter nucleic acid extraction liquid passage, enter rinsing liquid passage, enter EDTA SDS passage, enter nuclease passage, enter cell pyrolysis liquid passage to be connected with convergence passage respectively with the right-hand member entering cell suspension passage,
Converge the fore-and-aft direction that passage is positioned at the middle part of described fluid passage layer upper surface, the channel section converged on passage between the right-hand member entering EDTA SDS passage and the right-hand member entering nuclease passage is connected with the entrance point of reaction channel, reaction channel is positioned at the right side of fluid passage layer upper surface, its axis is positioned at left and right directions, reaction channel from left to right successively with the import of row's cell pyrolysis liquid passage, the outlet of microballoon admission passage is connected with the import of row's reactant liquor passage, row's cell pyrolysis liquid passage, microballoon admission passage and row's reactant liquor passage are positioned at the fore-and-aft direction on the right side of fluid passage layer upper surface, the outlet of row's cell pyrolysis liquid passage, the import of microballoon admission passage and the outlet of row's reactant liquor passage are positioned at front side or the rear side of fluid passage layer lower surface, the right-hand member of described reaction channel is provided with the DNA penetrating described fluid passage layer downwards and collects mouth,
The left side of described control channel layer lower surface is provided with in left-right direction from front to back successively into nucleic acid extraction liquid control channel, enter rinsing liquid control channel, enter EDTA SDS control channel, enter nuclease control channel, enter cell pyrolysis liquid control channel and enter cell suspension control channel, enter nucleic acid extraction liquid control channel, enter rinsing liquid control channel, enter EDTA SDS control channel, enter nuclease control channel, enter cell pyrolysis liquid control channel and be respectively equipped with the outer end entering cell suspension control channel the air inlet upwards penetrating described control channel layer, enter nucleic acid extraction liquid control channel to pass from the top entering nucleic acid extraction liquid passage, enter rinsing liquid control channel to pass from the top entering rinsing liquid passage, enter EDTA SDS control channel to pass from the top entering EDTA SDS passage, enter nuclease control channel to pass from the top entering nuclease passage, enter cell pyrolysis liquid control channel to pass from the top entering cell pyrolysis liquid passage, enter cell suspension control channel to pass from the top entering cell suspension passage,
The right side of the described control channel layer lower surface row's of being provided with cell pyrolysis liquid control channel successively from left to right, microballoon enters control channel and row's reactant liquor control channel, row's cell pyrolysis liquid control channel, microballoon enters control channel and row's reactant liquor control channel is arranged respectively along the longitudinal direction, row's cell pyrolysis liquid control channel, the outer end that microballoon enters control channel and row's reactant liquor control channel is respectively equipped with the air inlet upwards penetrating described control channel layer, row's cell pyrolysis liquid control channel is passed from the top of row's cell pyrolysis liquid passage, microballoon enters control channel and passes from the top of microballoon admission passage, row's reactant liquor control channel is passed from the top of row's reactant liquor passage,
The lower surface of described control channel layer is also provided with left process control channel, middle process control channel and right process control channel, left process control channel, the outer end of middle process control channel and right process control channel is respectively equipped with the air inlet upwards penetrating described control channel layer, pass above the channel section of left process control channel from described reaction channel between the import being positioned at row's cell pyrolysis liquid passage and the outlet of microballoon admission passage, described middle process control channel is passed being positioned at above the channel section between the outlet of microballoon admission passage and the import of described row's reactant liquor control channel from described reaction channel, described left process control channel is passed from described reaction channel above channel section is collected between mouth in the import and described DNA that are positioned at row's reactant liquor control channel.
The nucleic acid extraction chip of extraction gram-positive bacteria of the present invention and Gram-negative bacteria nucleic acid, wherein said enter the inner end of nucleic acid extraction liquid control channel pass from the top entering nucleic acid extraction liquid passage right-hand member, described enter rinsing liquid control channel inner end from described enter the top of rinsing liquid passage right-hand member pass, the inner end of the described EDTA of entering SDS control channel is passed from the top of the described EDTA of entering SDS passage right-hand member, described enter the inner end of nuclease control channel pass from the top entering nuclease passage right-hand member, described enter cell pyrolysis liquid control channel inner end from described enter the top of cell pyrolysis liquid passage right-hand member pass, the inner end entering cell suspension control channel is passed from the top entering cell suspension passage right-hand member, the inner end of described row's cell pyrolysis liquid control channel is passed from the top of row's cell pyrolysis liquid channel middle, the top of inner end in the middle part of microballoon admission passage that described microballoon enters control channel is passed, and the inner end of described row's reactant liquor control channel is passed from the top of described row's reactant liquor channel middle.
The nucleic acid extraction chip of extraction gram-positive bacteria of the present invention and Gram-negative bacteria nucleic acid, the thickness of wherein said control channel layer is 3-5 millimeter, the thickness of described elastic film layer is 50 microns, the thickness of described fluid passage layer is 150 microns, described enter nucleic acid extraction liquid passage, enter rinsing liquid passage, enter EDTA SDS passage, enter nuclease passage, enter cell pyrolysis liquid passage, enter cell suspension passage, converge passage, row's cell pyrolysis liquid passage, microballoon admission passage, the height of row's reactant liquor passage is all 100 microns, width is all 300 microns, length is all 5 millimeters, the height of described each control channel is all 100 microns, width is all 300 microns.
The nucleic acid extraction chip of extraction gram-positive bacteria of the present invention and Gram-negative bacteria nucleic acid, wherein said elastic film layer is dimethyl silicone polymer (PDMS).
Compared with prior art, beneficial effect of the present invention is:
1) under the nucleic acid extraction chip that the present invention extracts gram-positive bacteria and Gram-negative bacteria nucleic acid can be used in being not easy to the environment of artificial direct control, or the nucleic acid extraction be not suitable in the sample of artificial directly contact, also human cost can be saved, the development of related industry can be promoted, because each passage is controlled separately, regulation and control flexibly, therefore can process complex sample.
2) the present invention extracts the nucleic acid extraction chip of gram-positive bacteria and Gram-negative bacteria nucleic acid owing to not needing through centrifugal treating, be convenient to automation mechanized operation, simple, convenient, detect rapidly, effect is objective, can be scientific research, medical diagnosis on disease, genetic test, forensic identification etc. and provides technical support.
Below in conjunction with accompanying drawing, technical scheme of the present invention is described further.
Accompanying drawing explanation
Fig. 1 is the structural principle schematic diagram of the embodiment of the nucleic acid extraction chip of extraction gram-positive bacteria of the present invention and Gram-negative bacteria nucleic acid.
Detailed description of the invention
As shown in Figure 1, the present invention extracts the nucleic acid extraction chip of gram-positive bacteria and Gram-negative bacteria nucleic acid, comprises control channel layer, elastic film layer and fluid passage layer, the lower surface of control channel layer and the upper surface of elastic film layer are close to setting, the lower surface of elastic film layer and the upper surface of fluid passage layer are close to setting, and the left side of fluid passage layer upper surface is provided with in left-right direction from front to back successively into nucleic acid extraction liquid passage 6, enter rinsing liquid passage 5, enter EDTA SDS passage 4, enter nuclease passage 3, enter cell pyrolysis liquid passage 2 and enter cell suspension passage 1, entering nucleic acid extraction liquid passage 6, enter rinsing liquid passage 5, enter EDTA SDS passage 4, enter nuclease passage 3, enter cell pyrolysis liquid passage 2 and be respectively equipped with the left end entering cell suspension passage 1 inlet penetrating fluid passage layer downwards, enter nucleic acid extraction liquid passage 6, enter rinsing liquid passage 5, enter EDTA SDS passage 4, enter nuclease passage 3, enter cell pyrolysis liquid passage 2 to be connected with convergence passage 7 respectively with the right-hand member entering cell suspension passage 1,
Converge the fore-and-aft direction that passage 7 is positioned at the middle part of fluid passage layer upper surface, the channel section converged on passage 7 between the right-hand member entering EDTA SDS passage 4 and the right-hand member entering nuclease passage 3 is connected with the entrance point of reaction channel 8, reaction channel 8 is positioned at the right side of fluid passage layer upper surface, its axis is positioned at left and right directions, reaction channel 8 from left to right successively with the import of row's cell pyrolysis liquid passage 9, the outlet of microballoon admission passage 10 is connected with the import of row's reactant liquor passage 11, row's cell pyrolysis liquid passage 9, microballoon admission passage 10 and row's reactant liquor passage 11 are positioned at the fore-and-aft direction on the right side of fluid passage layer upper surface, the outlet of row's cell pyrolysis liquid passage 9, the import of microballoon admission passage 10 and the outlet of row's reactant liquor passage 11 are positioned at front side or the rear side of fluid passage layer lower surface, the right-hand member of reaction channel 8 is provided with the DNA penetrating fluid passage layer downwards and collects mouth 12,
The left side of control channel layer lower surface is provided with in left-right direction from front to back successively into nucleic acid extraction liquid control channel 26, enter rinsing liquid control channel 25, enter EDTA SDS control channel 24, enter nuclease control channel 23, enter cell pyrolysis liquid control channel 22 and enter cell suspension control channel 21, enter nucleic acid extraction liquid control channel 26, enter rinsing liquid control channel 25, enter EDTA SDS control channel 24, enter nuclease control channel 23, enter cell pyrolysis liquid control channel 22 and be respectively equipped with the outer end entering cell suspension control channel 21 air inlet upwards penetrating control channel layer, enter nucleic acid extraction liquid control channel 26 to pass from the top entering nucleic acid extraction liquid passage 6, enter rinsing liquid control channel 25 to pass from the top entering rinsing liquid passage 5, enter EDTASDS control channel 24 to pass from the top entering EDTA SDS passage 4, enter nuclease control channel 23 to pass from the top entering nuclease passage 3, enter cell pyrolysis liquid control channel 22 to pass from the top entering cell pyrolysis liquid passage 2, enter cell suspension control channel 21 to pass from the top entering cell suspension passage 1,
The right side of the control channel layer lower surface row's of being provided with cell pyrolysis liquid control channel 29 successively from left to right, microballoon enters control channel 13 and row's reactant liquor control channel 14, row's cell pyrolysis liquid control channel 29, microballoon enters control channel 13 and row's reactant liquor control channel 14 is arranged respectively along the longitudinal direction, row's cell pyrolysis liquid control channel 29, the outer end that microballoon enters control channel 13 and row's reactant liquor control channel 14 is respectively equipped with the air inlet upwards penetrating control channel layer, row's cell pyrolysis liquid control channel 29 is passed from the top of row's cell pyrolysis liquid passage 9, microballoon enters control channel 13 and passes from the top of microballoon admission passage 10, row's reactant liquor control channel 14 is passed from the top of row's reactant liquor passage 11,
The lower surface of control channel layer is also provided with left process control channel 15, middle process control channel 16 and right process control channel 17, left process control channel 15, the outer end of middle process control channel 16 and right process control channel 17 is respectively equipped with the air inlet upwards penetrating control channel layer, pass above the channel section of left process control channel 15 from reaction channel 8 between the import being positioned at row's cell pyrolysis liquid passage 9 and the outlet of microballoon admission passage 10, middle process control channel 16 is passed being positioned at above the channel section between the outlet of microballoon admission passage 10 and the import of row's reactant liquor control channel 14 from reaction channel 8, left process control channel 15 is passed from reaction channel 8 above channel section is collected between mouth 12 in the import and DNA that are positioned at row's reactant liquor control channel 14.
As improvement of the present invention, above-mentioned enter the inner end of nucleic acid extraction liquid control channel 26 pass from the top entering nucleic acid extraction liquid passage 6 right-hand member, the inner end entering rinsing liquid control channel 25 is passed from the top entering rinsing liquid passage 5 right-hand member, the inner end entering EDTA SDS control channel 24 is passed from the top entering EDTA SDS passage 4 right-hand member, the inner end entering nuclease control channel 23 is passed from the top entering nuclease passage 3 right-hand member, the inner end entering cell pyrolysis liquid control channel 22 is passed from the top entering cell pyrolysis liquid passage 2 right-hand member, the inner end entering cell suspension control channel 21 is passed from the top entering cell suspension passage 1 right-hand member, the top of inner end in the middle part of row's cell pyrolysis liquid passage 9 of row's cell pyrolysis liquid control channel 29 is passed, the top of inner end in the middle part of microballoon admission passage 10 that microballoon enters control channel 13 is passed, and the top of inner end in the middle part of row's reactant liquor passage 11 of row's reactant liquor control channel 14 is passed.
As improvement of the present invention, the thickness of above-mentioned control channel layer is 3-5 millimeter, the thickness of elastic film layer is 50 microns, the thickness of fluid passage layer is 150 microns, enter nucleic acid extraction liquid passage 6, enter rinsing liquid passage 5, enter EDTA SDS passage 4, enter nuclease passage 3, enter cell pyrolysis liquid passage 2, enter cell suspension passage 1, converge passage 7, row's cell pyrolysis liquid passage 9, microballoon admission passage 10, the height of row's reactant liquor passage 11 is all 100 microns, width is all 300 microns, length is all 5 millimeters, the height of each control channel is all 100 microns, width is all 300 microns.
As improvement of the present invention, above-mentioned elastic film layer is dimethyl silicone polymer (PDMS).
The operation principle of the nucleic acid extraction chip of extraction gram-positive bacteria of the present invention and Gram-negative bacteria nucleic acid is as follows: if to control channel charged pressure gas, pressed gas is generally nitrogen, then can by the elastic membrane of compressing elastic film layer, make it that deformation occur and by the fluid passage got into correspondence position, oppress and close this fluid passage, make liquid not by.If the pressed gas in control channel is released, then elastic membrane recovers origin-location, and corresponding fluid passage is unimpeded.
The concrete use step of the nucleic acid extraction chip of extraction gram-positive bacteria of the present invention and Gram-negative bacteria nucleic acid is as follows:
One, the chip fluid passage integrated is connected different solution, keep all control channel to be inflated condition during beginning, all fluid passage gateways are closed by control channel;
Two, the PBS suspension that the silicon oxide pellets of 50 micron diameters is housed is injected reaction channel by microballoon admission passage 10, make bead be taken in the space of the reaction channel 8 before screen cloth as far as possible;
Three, by other pathway closures, open into cell suspension passage 1 and row's cell pyrolysis liquid passage 9, Buffer1 (20mM Tris-HCl will be suspended in, 2mM EDTA, pH8.0 and SDS and TritonX-100, be 0.01%SDS and 1.2%TritonX-100 for E.coli. be 1%SDS and 2.4%TritonX-10 for gram-positive bacteria) bacterium liquid by cell suspension passage 1 injection channel, make solution enter reaction channel region, surplus liquid is discharged by arranging cell pyrolysis liquid passage 9.
Four, by other pathway closures, open into cell pyrolysis liquid passage 2 and row's reactant liquor passage 11, pass through into cell pyrolysis liquid passage 2 injection channel by buffer2 (the 3M GuSCN containing 0.8mg/ml proteinase K), unnecessary liquid is discharged by arranging reactant liquor passage 11.DNA can separate out and be adsorbed on silicon dioxide microsphere by this process by cellular lysate;
Five, by other pathway closures, opening and collect mouth 12 into nuclease passage 3 and DNA, being injected by 70% ethanol by entering nuclease passage 3, surplus liquid is collected mouth 12 by DNA and is discharged, and this step can by other Impurity removals except DNA;
Six, by other pathway closures, open and collect mouth 12 into EDTA SDS passage 4 and DNA, injected by entering EDTA SDS passage 4 by 100% ethanol, surplus liquid is collected mouth 12 by DNA and is discharged, and this step can be removed impurity further and make DNA more closely be attached to microsphere surface;
Seven, by other pathway closures, open fluid passage and enter rinsing liquid passage 5 and DNA collection mouth 12, clean nitrogen being introduced from entering rinsing liquid passage 5, collecting mouth 12 from DNA and discharging, dried up by ethanol (about needing 10 minutes), this step is removed by ethanol;
Eight, by other pathway closures, open and collect mouth 12 into nucleic acid extraction liquid passage 6 and DNA, pure water is injected from entering nucleic acid extraction liquid passage 6, the DNA on silicon dioxide microsphere is eluted, to DNA collection mouth 12, DNA is being collected;
Nine, open successively fluid passage enter cell suspension passage 1, enter cell pyrolysis liquid passage 2, enter nuclease passage 3, enter EDTA SDS passage 4, enter rinsing liquid passage 5, enter nucleic acid extraction liquid passage 6, row's cell pyrolysis liquid passage 9, microballoon admission passage 10, liquid collects mouth 12 by arranging reactant liquor passage 11 to DNA, inject PBS and water successively, chip the various piece of passage cleaned up, so that may be used for testing next time;
Ten, the flow rate of liquid of above-mentioned whole process controls at 1000-1500ul/h.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite not departing from the technology of the present invention principle; can also make some improvement and modification, these improve and modification also should be considered as protection scope of the present invention.
Claims (3)
1. extract the nucleic acid extraction chip of gram-positive bacteria and Gram-negative bacteria nucleic acid, it is characterized in that: comprise control channel layer, elastic film layer and fluid passage layer, the lower surface of control channel layer and the upper surface of elastic film layer are close to setting, the lower surface of elastic film layer and the upper surface of described fluid passage layer are close to setting, and the left side of described fluid passage layer upper surface is provided with in left-right direction from front to back successively into nucleic acid extraction liquid passage (6), enter rinsing liquid passage (5), enter EDTA SDS passage (4), enter nuclease passage (3), enter cell pyrolysis liquid passage (2) and enter cell suspension passage (1), entering nucleic acid extraction liquid passage (6), enter rinsing liquid passage (5), enter EDTA SDS passage (4), enter nuclease passage (3), the left end entering cell pyrolysis liquid passage (2) and enter cell suspension passage (1) is respectively equipped with the inlet penetrating described fluid passage layer downwards, enters nucleic acid extraction liquid passage (6), enter rinsing liquid passage (5), enter EDTA SDS passage (4), enter nuclease passage (3), enter cell pyrolysis liquid passage (2) to be connected with convergence passage (7) respectively with the right-hand member entering cell suspension passage (1),
Converge the fore-and-aft direction that passage (7) is positioned at the middle part of described fluid passage layer upper surface, convergence passage (7) is positioned at and is connected with the entrance point of reaction channel (8) into the channel section between the right-hand member and the right-hand member entering nuclease passage (3) of EDTA SDS passage (4), reaction channel (8) is positioned at the right side of fluid passage layer upper surface, its axis is positioned at left and right directions, reaction channel (8) from left to right successively with row cell pyrolysis liquid passage (9) import, the outlet of microballoon admission passage (10) is connected with the import of row's reactant liquor passage (11), row's cell pyrolysis liquid passage (9), microballoon admission passage (10) and row's reactant liquor passage (11) are positioned at the fore-and-aft direction on the right side of fluid passage layer upper surface, the outlet of row's cell pyrolysis liquid passage (9), the import of microballoon admission passage (10) and the outlet of row's reactant liquor passage (11) are positioned at front side or the rear side of fluid passage layer lower surface, the right-hand member of described reaction channel (8) is provided with the DNA penetrating described fluid passage layer downwards and collects mouth (12),
The left side of described control channel layer lower surface is provided with in left-right direction from front to back successively into nucleic acid extraction liquid control channel (26), enter rinsing liquid control channel (25), enter EDTA SDS control channel (24), enter nuclease control channel (23), enter cell pyrolysis liquid control channel (22) and enter cell suspension control channel (21), enter nucleic acid extraction liquid control channel (26), enter rinsing liquid control channel (25), enter EDTA SDS control channel (24), enter nuclease control channel (23), the outer end entered cell pyrolysis liquid control channel (22) and enter cell suspension control channel (21) is respectively equipped with the air inlet upwards penetrating described control channel layer, enter nucleic acid extraction liquid control channel (26) to pass from the top entering nucleic acid extraction liquid passage (6), enter rinsing liquid control channel (25) to pass from the top entering rinsing liquid passage (5), enter EDTA SDS control channel (24) to pass from the top entering EDTA SDS passage (4), enter nuclease control channel (23) to pass from the top entering nuclease passage (3), enter cell pyrolysis liquid control channel (22) to pass from the top entering cell pyrolysis liquid passage (2), enter cell suspension control channel (21) to pass from the top entering cell suspension passage (1),
The right side of the described control channel layer lower surface row's of being provided with cell pyrolysis liquid control channel (29) successively from left to right, microballoon enters control channel (13) and row's reactant liquor control channel (14), row's cell pyrolysis liquid control channel (29), microballoon enters control channel (13) and row's reactant liquor control channel (14) arranges respectively along the longitudinal direction, row's cell pyrolysis liquid control channel (29), the outer end that microballoon enters control channel (13) and row's reactant liquor control channel (14) is respectively equipped with the air inlet upwards penetrating described control channel layer, row's cell pyrolysis liquid control channel (29) passes from the top of row's cell pyrolysis liquid passage (9), microballoon enters control channel (13) and passes from the top of microballoon admission passage (10), row's reactant liquor control channel (14) passes from the top of row's reactant liquor passage (11),
The lower surface of described control channel layer is also provided with left process control channel (15), middle process control channel (16) and right process control channel (17), left process control channel (15), the outer end of middle process control channel (16) and right process control channel (17) is respectively equipped with the air inlet upwards penetrating described control channel layer, left process control channel (15) is passed above described reaction channel (8) is positioned at the channel section between the import of row's cell pyrolysis liquid passage (9) and the outlet of microballoon admission passage (10), described middle process control channel (16) is passed above described reaction channel (8) is positioned at the channel section between the outlet of microballoon admission passage (10) and the import of described row's reactant liquor control channel (14), described left process control channel (15) is passed above described reaction channel (8) is positioned at the channel section between the import of row's reactant liquor control channel (14) and described DNA collection mouth (12),
Described enter the inner end of nucleic acid extraction liquid control channel (26) pass from the top entering nucleic acid extraction liquid passage (6) right-hand member, described enter rinsing liquid control channel (25) inner end from described enter the top of rinsing liquid passage (5) right-hand member pass, the inner end of the described EDTA of entering SDS control channel (24) is passed from the top of the described EDTA of entering SDS passage (4) right-hand member, described enter the inner end of nuclease control channel (23) pass from the top entering nuclease passage (3) right-hand member, described enter cell pyrolysis liquid control channel (22) inner end from described enter the top of cell pyrolysis liquid passage (2) right-hand member pass, the inner end entering cell suspension control channel (21) is passed from the top entering cell suspension passage (1) right-hand member, the inner end of described row's cell pyrolysis liquid control channel (29) is passed from the top at row's cell pyrolysis liquid passage (9) middle part, the inner end that described microballoon enters control channel (13) is passed from the top at microballoon admission passage (10) middle part, and the inner end of described row's reactant liquor control channel (14) is passed from the top at described row's reactant liquor passage (11) middle part.
2. according to the nucleic acid extraction chip of extraction gram-positive bacteria according to claim 1 and Gram-negative bacteria nucleic acid, it is characterized in that: the thickness of described control channel layer is 3-5 millimeter, the thickness of described elastic film layer is 50 microns, the thickness of described fluid passage layer is 150 microns, described enter nucleic acid extraction liquid passage (6), enter rinsing liquid passage (5), enter EDTA SDS passage (4), enter nuclease passage (3), enter cell pyrolysis liquid passage (2), enter cell suspension passage (1), converge passage (7), row's cell pyrolysis liquid passage (9), microballoon admission passage (10), the height of row's reactant liquor passage (11) is all 100 microns, width is all 300 microns, length is all 5 millimeters, the height of described each control channel is all 100 microns, width is all 300 microns.
3., according to the nucleic acid extraction chip of extraction gram-positive bacteria according to claim 2 and Gram-negative bacteria nucleic acid, it is characterized in that: described elastic film layer is dimethyl silicone polymer (PDMS).
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| CN111304058B (en) * | 2020-03-26 | 2023-11-28 | 河南科技大学 | Microfluidic chip for detecting gram-negative bacteria genes and detection method |
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| JP4445611B2 (en) * | 1999-09-24 | 2010-04-07 | オリンパス株式会社 | Equipment for polymorphism analysis of repetitive sequences |
| US20060194207A1 (en) * | 2003-07-30 | 2006-08-31 | Riken | Kit for detecting nucleic acids |
| KR20120063162A (en) * | 2010-12-07 | 2012-06-15 | 삼성전자주식회사 | Gene analysis apparatus and method of analyzing gene using the same |
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| CN102115711A (en) * | 2010-01-06 | 2011-07-06 | 深圳先进技术研究院 | Micro-flow control chip and nucleic acid extracting and purifying method |
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| CN103484354A (en) | 2014-01-01 |
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