CN103484354A - 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 that extracts gram-positive microorganism 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 several different methods and have extracted nucleic acid.But wherein great majority relate to centrifugally, this is unfavorable for realizing microminiaturization and the automatization of nucleic acid extraction.
Summary of the invention
The object of the present invention is to provide a kind of the needs through centrifugal treating, be convenient to automated operation, simple, convenient, detect rapidly, effect is objective, can be the nucleic acid extraction chip based on silica bead method extraction gram-positive microorganism and Gram-negative bacteria nucleic acid that scientific research, medical diagnosis on disease, gene test, forensic identification etc. provide technical support.
The present invention can extract the nucleic acid extraction chip of gram-positive microorganism and Gram-negative bacteria nucleic acid, comprises the 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 upper surface of the lower surface of elastic film layer and described fluid passage layer is close to setting, and the left side of described fluid passage layer upper surface is provided with into nucleic acid extraction liquid passage from front to back successively along left and right directions, advance the rinsing liquid passage, advance EDTA SDS passage, advance the nuclease passage, advance the cell pyrolysis liquid passage and advance the cell suspension passage, advancing nucleic acid extraction liquid passage, advance the rinsing liquid passage, advance EDTA SDS passage, advance the nuclease passage, advance the cell pyrolysis liquid passage and be respectively equipped with the left end that advances the cell suspension passage fluid inlet that penetrates described fluid passage layer downwards, advance nucleic acid extraction liquid passage, advance the rinsing liquid passage, advance EDTA SDS passage, advance the nuclease passage, advance cell pyrolysis liquid passage and the right-hand member that advances the cell suspension passage respectively with converge passage and be connected,
Converge passage and be positioned at the fore-and-aft direction at the middle part of described fluid passage layer upper surface, converge on passage at the right-hand member that advances EDTA SDS passage and the channel section that advances between the right-hand member of nuclease passage and be connected with the entrance end 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 reaction solution passage, row's cell pyrolysis liquid passage, microballoon admission passage and row's reaction solution 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 outlet of the import of microballoon admission passage and row's reaction solution passage is 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 that penetrates described fluid passage layer downwards and collects mouth,
The left side of described control channel layer lower surface is provided with into nucleic acid extraction hydraulic control passage processed from front to back successively along left and right directions, advance rinsing liquid control channel, advance EDTA SDS control channel, advance nuclease control channel, advance cell pyrolysis liquid control channel and advance cell suspension control channel, advance nucleic acid extraction hydraulic control passage processed, advance rinsing liquid control channel, advance EDTA SDS control channel, advance nuclease control channel, advance cell pyrolysis liquid control channel and be respectively equipped with the outer end of advancing cell suspension control channel the inlet mouth that upwards penetrates described control channel layer, advancing nucleic acid extraction hydraulic control passage processed passes from the top of advancing nucleic acid extraction liquid passage, advancing rinsing liquid control channel passes from the top of advancing the rinsing liquid passage, advancing EDTA SDS control channel passes from the top of advancing EDTA SDS passage, advancing nuclease control channel passes from the top of advancing the nuclease passage, advancing cell pyrolysis liquid control channel passes from the top of advancing the cell pyrolysis liquid passage, advancing cell suspension control channel passes from the top of advancing the cell suspension passage,
The right side of described control channel layer lower surface is the row's of being provided with cell pyrolysis liquid control channel successively from left to right, microballoon enters control channel and row's reaction solution control channel, row's cell pyrolysis liquid control channel, microballoon enters control channel and row's reaction solution control channel is arranged along fore-and-aft direction respectively, row's cell pyrolysis liquid control channel, the outer end that microballoon enters control channel and row's reaction solution control channel is respectively equipped with the inlet mouth that upwards penetrates 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 reaction solution control channel is passed from the top of row's reaction solution passage,
The lower surface of described control channel layer also is provided with left processing controls passage, middle processing controls passage and right processing controls passage, left processing controls passage, the outer end of middle processing controls passage and right processing controls passage is respectively equipped with the inlet mouth that upwards penetrates described control channel layer, left processing controls passage passes from described reaction channel above the channel section between the outlet of the import that is positioned at row's cell pyrolysis liquid passage and microballoon admission passage, described middle processing controls passage passes from described reaction channel above the channel section between the import of the outlet that is positioned at the microballoon admission passage and described row's reaction solution control channel, described left processing controls passage passes from described reaction channel above the channel section between the import that is positioned at row's reaction solution control channel and described DNA collection mouth.
The nucleic acid extraction chip that extracts gram-positive microorganism and Gram-negative bacteria nucleic acid of the present invention, the wherein said inner end of advancing nucleic acid extraction hydraulic control passage processed is passed from the top of advancing nucleic acid extraction liquid passage right-hand member, the described inner end of advancing rinsing liquid control channel is passed from the described top of advancing rinsing liquid passage right-hand member, the inner end of the described EDTA of advancing SDS control channel is passed from the top of the described EDTA of advancing SDS passage right-hand member, the described inner end of advancing nuclease control channel is passed from the top of advancing nuclease passage right-hand member, the described inner end of advancing cell pyrolysis liquid control channel is passed from the described top of advancing cell pyrolysis liquid passage right-hand member, entering the inner end of cell suspension control channel passes from the top of advancing 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, described microballoon enters the inner end of control channel and passes from the top at microballoon admission passage middle part, and the inner end of described row's reaction solution control channel is passed from the top of described row's reaction solution channel middle.
The nucleic acid extraction chip that extracts gram-positive microorganism and Gram-negative bacteria nucleic acid of the present invention, the thickness of wherein said control channel layer is the 3-5 millimeter, the thickness of described elastic film layer is 50 microns, the thickness of described fluid passage layer is 150 microns, describedly advance nucleic acid extraction liquid passage, advance the rinsing liquid passage, advance EDTA SDS passage, advance the nuclease passage, advance the cell pyrolysis liquid passage, advance the cell suspension passage, converge passage, row's cell pyrolysis liquid passage, the microballoon admission passage, the height of row's reaction solution 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 that extracts gram-positive microorganism and Gram-negative bacteria nucleic acid of the present invention, wherein said elastic film layer is polydimethylsiloxane (PDMS).
Compared with prior art, beneficial effect of the present invention is:
1) the nucleic acid extraction chip that the present invention can extract gram-positive microorganism and Gram-negative bacteria nucleic acid can be used under the environment that is not easy to artificial direct control, perhaps be not suitable for the nucleic acid extraction in the artificial sample directly contacted, also can save human cost, can promote the development of related industries, because each passage is controlled separately, regulation and control are flexible, therefore can process complex sample.
2) the present invention can extract the nucleic acid extraction chip of gram-positive microorganism and Gram-negative bacteria nucleic acid owing to not needing through centrifugal treating, be convenient to automated operation, simple, convenient, detect rapidly, effect is objective, can be scientific research, medical diagnosis on disease, gene test, forensic identification etc. technical support is provided.
Below in conjunction with accompanying drawing, technical scheme of the present invention is described further.
The accompanying drawing explanation
Fig. 1 is the structural principle schematic diagram of the embodiment of the nucleic acid extraction chip that extracts gram-positive microorganism and Gram-negative bacteria nucleic acid of the present invention.
Embodiment
As shown in Figure 1, the present invention can extract the nucleic acid extraction chip of gram-positive microorganism and Gram-negative bacteria nucleic acid, comprises the 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 upper surface of the lower surface of elastic film layer and fluid passage layer is close to setting, and the left side of fluid passage layer upper surface is provided with into nucleic acid extraction liquid passage 6 from front to back successively along left and right directions, advance rinsing liquid passage 5, advance EDTASDS passage 4, advance nuclease passage 3, advance cell pyrolysis liquid passage 2 and advance cell suspension passage 1, advancing nucleic acid extraction liquid passage 6, advance rinsing liquid passage 5, advance EDTA SDS passage 4, advance nuclease passage 3, advance cell pyrolysis liquid passage 2 and be respectively equipped with the left end that advances cell suspension passage 1 fluid inlet that penetrates the fluid passage layer downwards, advance nucleic acid extraction liquid passage 6, advance rinsing liquid passage 5, advance EDTA SDS passage 4, advance nuclease passage 3, advance cell pyrolysis liquid passage 2 and the right-hand member that advances cell suspension passage 1 respectively with converge passage 7 and be connected,
Converge passage 7 and be positioned at the fore-and-aft direction at the middle part of fluid passage layer upper surface, converge on passage 7 at the right-hand member that advances EDTA SDS passage 4 and the channel section that advances between the right-hand member of nuclease passage 3 and be connected with the entrance end 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 reaction solution passage 11, row's cell pyrolysis liquid passage 9, microballoon admission passage 10 and row's reaction solution 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 outlet of the import of microballoon admission passage 10 and row's reaction solution passage 11 is 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 that penetrates the fluid passage layer downwards and collects mouth 12,
The left side of control channel layer lower surface is provided with into nucleic acid extraction hydraulic control passage 26 processed from front to back successively along left and right directions, advance rinsing liquid control channel 25, advance EDTA SDS control channel 24, advance nuclease control channel 23, advance cell pyrolysis liquid control channel 22 and advance cell suspension control channel 21, advance nucleic acid extraction hydraulic control passage 26 processed, advance rinsing liquid control channel 25, advance EDTA SDS control channel 24, advance nuclease control channel 23, advance cell pyrolysis liquid control channel 22 and be respectively equipped with the outer end of advancing cell suspension control channel 21 inlet mouth that upwards penetrates the control channel layer, advancing nucleic acid extraction hydraulic control passage 26 processed passes from the top of advancing nucleic acid extraction liquid passage 6, advancing rinsing liquid control channel 25 passes from the top of advancing rinsing liquid passage 5, advancing EDTASDS control channel 24 passes from the top of advancing EDTA SDS passage 4, advancing nuclease control channel 23 passes from the top of advancing nuclease passage 3, advancing cell pyrolysis liquid control channel 22 passes from the top of advancing cell pyrolysis liquid passage 2, advancing cell suspension control channel 21 passes from the top of advancing cell suspension passage 1,
The right side of control channel layer lower surface is the 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 reaction solution control channel 14, row's cell pyrolysis liquid control channel 29, microballoon enters control channel 13 and row's reaction solution control channel 14 is arranged along fore-and-aft direction respectively, row's cell pyrolysis liquid control channel 29, the outer end that microballoon enters control channel 13 and row's reaction solution control channel 14 is respectively equipped with the inlet mouth that upwards penetrates the 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 reaction solution control channel 14 is passed from the top of row's reaction solution passage 11,
The lower surface of control channel layer also is provided with left processing controls passage 15, middle processing controls passage 16 and right processing controls passage 17, left processing controls passage 15, the outer end of middle processing controls passage 16 and right processing controls passage 17 is respectively equipped with the inlet mouth that upwards penetrates the control channel layer, left processing controls passage 15 passes from reaction channel 8 above the channel section between the outlet of the import that is positioned at row's cell pyrolysis liquid passage 9 and microballoon admission passage 10, middle processing controls passage 16 passes from reaction channel 8 above the channel section between the import of the outlet that is positioned at microballoon admission passage 10 and row's reaction solution control channel 14, left processing controls passage 15 passes from reaction channel 8 above the channel section between the import that is positioned at row's reaction solution control channel 14 and DNA collection mouth 12.
As improvement of the present invention, the above-mentioned inner end of advancing nucleic acid extraction hydraulic control passage 26 processed is passed from the top of advancing nucleic acid extraction liquid passage 6 right-hand members, entering the inner end of rinsing liquid control channel 25 passes from the top of advancing rinsing liquid passage 5 right-hand members, entering the inner end of EDTA SDS control channel 24 passes from the top of advancing EDTA SDS passage 4 right-hand members, entering the inner end of nuclease control channel 23 passes from the top of advancing nuclease passage 3 right-hand members, entering the inner end of cell pyrolysis liquid control channel 22 passes from the top of advancing cell pyrolysis liquid passage 2 right-hand members, entering the inner end of cell suspension control channel 21 passes from the top of advancing cell suspension passage 1 right-hand member, the inner end of row's cell pyrolysis liquid control channel 29 is passed from the top at row cell pyrolysis liquid passage 9 middle parts, microballoon enters the inner end of control channel 13 and passes from the top at microballoon admission passage 10 middle parts, and the inner end of row's reaction solution control channel 14 is passed from the top at row reaction solution passage 11 middle parts.
As improvement of the present invention, the thickness of above-mentioned control channel layer is the 3-5 millimeter, the thickness of elastic film layer is 50 microns, the thickness of fluid passage layer is 150 microns, advance nucleic acid extraction liquid passage 6, advance rinsing liquid passage 5, advance EDTA SDS passage 4, advance nuclease passage 3, advance cell pyrolysis liquid passage 2, advance cell suspension passage 1, converge passage 7, row's cell pyrolysis liquid passage 9, microballoon admission passage 10, the height of row's reaction solution 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 polydimethylsiloxane (PDMS).
The principle of work of the nucleic acid extraction chip that extracts gram-positive microorganism and Gram-negative bacteria nucleic acid of the present invention is as follows: if to control channel charged pressure gas, pressed gas is generally nitrogen, can be by the elastica of compressing elastic film layer, make it deformation occur and got into the fluid passage of correspondence position, oppress and seal this fluid passage, liquid can not be passed through.If the pressed gas in control channel is released, elastica is recovered home position, and corresponding fluid passage is unimpeded.
The concrete use step of the nucleic acid extraction chip that extracts gram-positive microorganism and Gram-negative bacteria nucleic acid of the present invention is as follows:
One, the chip fluid passage of having integrated is connected to different solution, during beginning, keep all control channel to be inflated condition, control channel is by all fluid passage gangway sealings;
The PBS suspension that two, the silicon-dioxide bead of 50 micron diameters will be housed injects reaction channel by microballoon admission passage 10, makes bead that the space of the reaction channel 8 before screen cloth is taken as far as possible;
Three, by other pathway closures, open into cell suspension passage 1 and row's cell pyrolysis liquid passage 9, to be suspended in Bufferl (20mM Tris-HCl, 2mM EDTA, pH8.0 and SDS and TritonX-100, for E.coli., be that 0.01%SDS and 1.2%TritonX-100 are 1%SDS and 2.4%TritonX-10 for gram-positive microorganism) bacterium liquid by cell suspension passage 1 injection channel, make solution enter the reaction channel zone, unnecessary liquid is discharged by row's cell pyrolysis liquid passage 9.
Four, by other pathway closures, open into cell pyrolysis liquid passage 2 and row's reaction solution passage 11, by buffer2 (the 3M GuSCN that contains 0.8mg/ml proteinase K), by entering cell pyrolysis liquid passage 2 injection channels, unnecessary liquid is discharged by row's reaction solution passage 11.This process can be separated out and be adsorbed on silicon dioxide microsphere by cellular lysate and by DNA;
Five, by other pathway closures, open into nuclease passage 3 and DNA and collect mouth 12,70% ethanol is injected by advancing nuclease passage 3, unnecessary liquid is collected mouth 12 by DNA and is discharged, and this step can be by other Impurity removals except DNA;
Six, by other pathway closures, open into EDTA SDS passage 4 and DNA and collect mouth 12,100% ethanol is injected by advancing EDTA SDS passage 4, and unnecessary liquid is collected mouthful 12 discharges by DNA, and this step can further be removed impurity and make DNA more closely be attached to microsphere surface;
Seven, by other pathway closures, open fluid passage and advance rinsing liquid passage 5 and DNA collection mouth 12, clean nitrogen is introduced from advancing rinsing liquid passage 5, collect mouthful 12 discharges from DNA, ethanol is dried up to (approximately needing 10 minutes), this step is that ethanol is removed;
Eight, by other pathway closures, open into nucleic acid extraction liquid passage 6 and DNA and collect mouth 12, pure water is injected from advancing 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 advance cell suspension passage 1, advance cell pyrolysis liquid passage 2,, advance nuclease passage 3, advance EDTA SDS passage 4, advance rinsing liquid passage 5, advance nucleic acid extraction liquid passage 6, row's cell pyrolysis liquid passage 9, microballoon admission passage 10, liquid collect mouthfuls 12 by row's reaction solution passage 11 to DNA, inject successively PBS and water, the various piece of passage is cleaned up, so that chip can be for experiment next time;
Ten, the flow rate of liquid of above-mentioned whole process is controlled 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 that does not break away 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 (4)
1. can extract the nucleic acid extraction chip of gram-positive microorganism and Gram-negative bacteria nucleic acid, it is characterized in that: comprise the 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 upper surface of the lower surface of elastic film layer and described fluid passage layer is close to setting, and the left side of described fluid passage layer upper surface is provided with into nucleic acid extraction liquid passage (6) from front to back successively along left and right directions, advance rinsing liquid passage (5), advance EDTA SDS passage (4), advance nuclease passage (3), advance cell pyrolysis liquid passage (2) and advance cell suspension passage (1), advancing nucleic acid extraction liquid passage (6), advance rinsing liquid passage (5), advance EDTA SDS passage (4), advance nuclease passage (3), the left end that advances cell pyrolysis liquid passage (2) and advance cell suspension passage (1) is respectively equipped with the fluid inlet that penetrates described fluid passage layer downwards, advances nucleic acid extraction liquid passage (6), advance rinsing liquid passage (5), advance EDTA SDS passage (4), advance nuclease passage (3), advance cell pyrolysis liquid passage (2) and the right-hand member that advances cell suspension passage (1) respectively with converge passage (7) and be connected,
Converge passage (7) and be positioned at the fore-and-aft direction at the middle part of described fluid passage layer upper surface, converge on passage (7) to be positioned at into the right-hand member of EDTA SDS passage (4) and the channel section that advances between the right-hand member of nuclease passage (3) and be connected with the entrance end 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 row the cell pyrolysis liquid passage (9) import, the outlet of microballoon admission passage (10) is connected with the import of row's reaction solution passage (11), row's cell pyrolysis liquid passage (9), microballoon admission passage (10) and row's reaction solution 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 outlet of the import of microballoon admission passage (10) and row's reaction solution passage (11) is 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 that penetrates described fluid passage layer downwards and collects mouthful (12),
The left side of described control channel layer lower surface is provided with into nucleic acid extraction hydraulic control passage processed (26) from front to back successively along left and right directions, advance rinsing liquid control channel (25), advance EDTA SDS control channel (24), advance nuclease control channel (23), advance cell pyrolysis liquid control channel (22) and advance cell suspension control channel (21), advance nucleic acid extraction hydraulic control passage processed (26), advance rinsing liquid control channel (25), advance EDTA SDS control channel (24), advance nuclease control channel (23), the outer end of advancing cell pyrolysis liquid control channel (22) and advancing cell suspension control channel (21) is respectively equipped with the inlet mouth that upwards penetrates described control channel layer, advancing nucleic acid extraction hydraulic control passage processed (26) passes from the top of advancing nucleic acid extraction liquid passage (6), advancing rinsing liquid control channel (25) passes from the top of advancing rinsing liquid passage (5), advancing EDTA SDS control channel (24) passes from the top of advancing EDTA SDS passage (4), advancing nuclease control channel (23) passes from the top of advancing nuclease passage (3), advancing cell pyrolysis liquid control channel (22) passes from the top of advancing cell pyrolysis liquid passage (2), advancing cell suspension control channel (21) passes from the top of advancing cell suspension passage (1),
The right side of described control channel layer lower surface is the 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 reaction solution control channel (14), row's cell pyrolysis liquid control channel (29), microballoon enters control channel (13) and row's reaction solution control channel (14) is arranged along fore-and-aft direction respectively, row's cell pyrolysis liquid control channel (29), the outer end that microballoon enters control channel (13) and row's reaction solution control channel (14) is respectively equipped with the inlet mouth that upwards penetrates described 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 reaction solution control channel (14) is passed from the top of row's reaction solution passage (11),
The lower surface of described control channel layer also is provided with left processing controls passage (15), middle processing controls passage (16) and right processing controls passage (17), left processing controls passage (15), the outer end of middle processing controls passage (16) and right processing controls passage (17) is respectively equipped with the inlet mouth that upwards penetrates described control channel layer, import and the top of the channel section between the outlet of microballoon admission passage (10) that left processing controls passage (15) is positioned at row's cell pyrolysis liquid passage (9) from described reaction channel (8) are passed, the channel section top that described middle processing controls passage (16) is positioned between the import of the outlet of microballoon admission passage (10) and described row's reaction solution control channel (14) from described reaction channel (8) is passed, described left processing controls passage (15) is positioned at the import of row's reaction solution control channel (14) and channel section top that described DNA collects between mouthful (12) is passed from described reaction channel (8).
2. according to the nucleic acid extraction chip that extracts gram-positive microorganism and Gram-negative bacteria nucleic acid claimed in claim 1, it is characterized in that: the described inner end of advancing nucleic acid extraction hydraulic control passage processed (26) is passed from the top of advancing nucleic acid extraction liquid passage (6) right-hand member, the described inner end of advancing rinsing liquid control channel (25) is passed from the described top of advancing rinsing liquid passage (5) right-hand member, the inner end of the described EDTA of advancing SDS control channel (24) is passed from the top of the described EDTA of advancing SDS passage (4) right-hand member, the described inner end of advancing nuclease control channel (23) is passed from the top of advancing nuclease passage (3) right-hand member, the described inner end of advancing cell pyrolysis liquid control channel (22) is passed from the described top of advancing cell pyrolysis liquid passage (2) right-hand member, entering the inner end of cell suspension control channel (21) passes from the top of advancing 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, described microballoon enters the inner end of control channel (13) and passes from the top at microballoon admission passage (10) middle part, and the inner end of described row's reaction solution control channel (14) is passed from the top at described row's reaction solution passage (11) middle part.
3. according to the nucleic acid extraction chip that extracts gram-positive microorganism and Gram-negative bacteria nucleic acid claimed in claim 2, it is characterized in that: the thickness of described control channel layer is the 3-5 millimeter, the thickness of described elastic film layer is 50 microns, the thickness of described fluid passage layer is 150 microns, describedly advance nucleic acid extraction liquid passage (6), advance rinsing liquid passage (5), advance EDTA SDS passage (4), advance nuclease passage (3), advance cell pyrolysis liquid passage (2), advance cell suspension passage (1), converge passage (7), row's cell pyrolysis liquid passage (9), microballoon admission passage (10), the height of row's reaction solution 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.
4. according to the nucleic acid extraction chip that extracts gram-positive microorganism and Gram-negative bacteria nucleic acid claimed in claim 3, it is characterized in that: described elastic film layer is polydimethylsiloxane (PDMS).
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107828633A (en) * | 2017-12-14 | 2018-03-23 | 苏州天隆生物科技有限公司 | A kind of nucleic acid extraction and reagent device, reagent and the method for detection |
| CN111304058A (en) * | 2020-03-26 | 2020-06-19 | 河南科技大学 | Microfluidic chip for detecting gram-negative bacteria gene and detection method |
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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 |
| US20120141999A1 (en) * | 2010-12-07 | 2012-06-07 | Samsung Electronics Co., Ltd. | Gene analysis apparatus and gene analysis method using the same |
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2013
- 2013-07-16 CN CN201310296807.XA patent/CN103484354B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001086993A (en) * | 1999-09-24 | 2001-04-03 | Olympus Optical Co Ltd | Device for analyzing polymorphism of reiterative sequence |
| US20060194207A1 (en) * | 2003-07-30 | 2006-08-31 | Riken | Kit for detecting nucleic acids |
| 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 |
| US20120141999A1 (en) * | 2010-12-07 | 2012-06-07 | Samsung Electronics Co., Ltd. | Gene analysis apparatus and gene analysis method using the same |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107828633A (en) * | 2017-12-14 | 2018-03-23 | 苏州天隆生物科技有限公司 | A kind of nucleic acid extraction and reagent device, reagent and the method for detection |
| CN107828633B (en) * | 2017-12-14 | 2021-04-27 | 苏州天隆生物科技有限公司 | Reagent device, reagent and method for extracting and detecting nucleic acid |
| CN111304058A (en) * | 2020-03-26 | 2020-06-19 | 河南科技大学 | Microfluidic chip for detecting gram-negative bacteria gene and detection method |
| 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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