CN103642674A - In situ polymerase chain reaction plate - Google Patents
In situ polymerase chain reaction plate Download PDFInfo
- Publication number
- CN103642674A CN103642674A CN201310674274.4A CN201310674274A CN103642674A CN 103642674 A CN103642674 A CN 103642674A CN 201310674274 A CN201310674274 A CN 201310674274A CN 103642674 A CN103642674 A CN 103642674A
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- Prior art keywords
- chain reaction
- situ polymerization
- polymerization enzyme
- enzyme chain
- plate
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0689—Sealing
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Hematology (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention provides an in situ polymerase chain reaction plate. The in situ polymerase chain reaction plate comprises an upper plate and a lower plate, wherein a sealing strip is arranged on the upper plate, the upper plate and the lower plate are connected by the sealing strip to form a cavity provided with an opening and used for accommodating a polymerase chain reaction reagent, and the in situ polymerase chain reaction plate is also provided with a sealing cover used for sealing the cavity. When the in situ polymerase chain reaction experiment is performed, the in situ polymerase chain reaction reagent is filled in the cavity and is sealed by using the sealing cover, thus ensuring the effectiveness of sealing in the in situ polymerase chain reaction process. The in situ polymerase chain reaction reagent does not evaporate and reacts completely, and experimental products do not pollute the environment, so that the success rate of the experiment is increased, thus ensuring diagnosis of specific genes and pathogens and reliably applying and popularizing the in situ polymerase chain reaction technology.
Description
Technical field
The present invention relates to polymerase chain reaction device, particularly relate to a kind of in-situ polymerization enzyme chain reaction plate.
Background technology
In-situ polymerization enzyme chain reaction technology successfully combines polymerase chain reaction technique and hybridization in situ technique, has kept the advantage of two technology to make up again deficiency separately.The sample to be checked of in-situ polymerization enzyme chain reaction technology is generally tissue slice or the cell on slide glass, and what first fixes through chemistry tissue slice or cell, to keep histiocytic good order and condition.Cytolemma and nuclear membrane all have certain permeability.When carrying out PCR amplification, various compositions (as primer, archaeal dna polymerase, Nucleotide etc.) all can enter in cell and nucleus in, take and be fixed in cell or endonuclear RNA or DNA are template, increase in position.The general molecule of product of amplification is larger, is difficult for through cytolemma, thus the original position of being retained in.In original like this cell, specific DNA or the RNA sequence of single copy or low copy are increased in position in a large number, and the product of amplification is just easy to be detected by hybridization in situ technique.Therefore, in-situ polymerization enzyme chain reaction technology can reflect kind, subspecies, the quantity of tested DNA or RNA, also reflects the kind of its place cell, thereby analyzes accurately or diagnose simultaneously.
Original position gene amplification is a kind of effective detection means, is widely used already, but requires high to its operative technique.But practice in owing to carrying out PCR amplification on slide glass, what seal use is traditional cover glass, not only operate very inconvenient, and can produce the problems such as gas leakage, evaporate to dryness, interference application, the bad searching of reaction conditions, thereby cause experiment success rate low, result is very unstable, has seriously restricted the reliability of in situ detection technology.
Summary of the invention
Based on this, be necessary for the problems referred to above, a kind of in-situ polymerization enzyme chain reaction plate when carrying out polymerase chain reaction experiment with low rate of evaporation, high success rate is provided.
A kind of in-situ polymerization enzyme chain reaction plate, comprise upper plate and lower plate, surface of described upper plate is provided with sealed strip, described upper plate and described lower plate by described sealed strip be connected to form have opening for holding the cavity of pcr reagent, described in-situ polymerization enzyme chain reaction plate also comprises sealing the sealing cover of described cavity.
In an embodiment, the surface relative with described upper plate of described lower plate is provided with the separation rete with net distribution that adopts bonding technique and protein cross technology to form therein.
In an embodiment, described upper plate adopts transparent material to make therein.
In an embodiment, the fluent material that described sealing cover is less than water by density forms therein.
In an embodiment, described fluent material is oil or whiteruss therein.
In an embodiment, described sealed strip is made by elaxtic seal therein.
Therein in an embodiment, described in-situ polymerization enzyme chain reaction plate is placed while using and sea line formation angle, described angle at 0 degree between 180 degree.
Therein in an embodiment, described lower plate is made by glass or polycarbonate or polymethylmethacrylate.
In an embodiment, on described in-situ polymerization enzyme chain reaction plate, also have deflector therein, described deflector is to be extended and form on the opening direction of described cavity by described lower plate.
Therein in an embodiment, that described sealed strip is while connecting upper plate and lower plate is U-shaped, V-type or C type.
Above-mentioned in-situ polymerization enzyme chain reaction plate comprises upper plate and lower plate, upper plate is provided with sealed strip, between upper plate and lower plate by sealed strip be connected to form have opening for holding the cavity of pcr reagent, and this in-situ polymerization enzyme chain reaction plate is also provided with the sealing cover in order to sealed cavity.When carrying out the experiment of in-situ polymerization enzyme chain reaction, in-situ polymerization enzyme chain reaction reagent is filled in this cavity and uses sealing cover to seal, guaranteed the validity sealing in in-situ polymerization enzyme chain reaction process.In-situ polymerization enzyme chain reaction reagent does not evaporate, and react completely, and experimental product does not pollute the environment, and has promoted the success ratio of experiment, thereby the diagnosis of specific gene and pathogenic agent is guaranteed, in-situ polymerization enzyme chain reaction technology is able to reliability application and popularization.
Accompanying drawing explanation
Fig. 1 is in-situ polymerization enzyme chain reaction plate vertical view;
Fig. 2 is the sectional view at in-situ polymerization enzyme chain reaction plate Y place;
Fig. 3 is the cut-away view of in-situ polymerization enzyme chain reaction plate;
Fig. 4 is lower plate schematic diagram;
Fig. 5 is upper plate schematic diagram.
Embodiment
Below in conjunction with accompanying drawing, preferred embodiment of the present invention is described in detail, thereby so that advantages and features of the invention can be easier to be it will be appreciated by those skilled in the art that, protection scope of the present invention is made to more explicit defining.
Please refer to Fig. 1, Fig. 2, Fig. 3 and Fig. 5, an embodiment of the invention provide a kind of in-situ polymerization enzyme chain reaction plate 100.This in-situ polymerization enzyme chain reaction plate 100 comprises upper plate 110, lower plate 120, and this upper plate 110 is provided with sealed strip 130.When carrying out the experiment of in-situ polymerization enzyme chain reaction, need first lower plate 120 to be carried out to pre-treatment, again upper plate 110 is connected by sealed strip 130 with through pretreated lower plate 120, and formation have opening for holding the cavity 140 of in-situ polymerization enzyme chain reaction reagent 150.Sealing bar 130 is arranged on a surface of upper plate 110 and is U-shaped, in other embodiments, can also be the shapes such as V-type, C type, as long as can meet the requirement of holding in-situ polymerization enzyme chain reaction reagent 150, is not specifically limited herein.Sealing bar 130 can be semiclosed sealing-ring, also can be seal gum, sealed strip 130 adopts elaxtic seal to make, in the present embodiment, adopt silicon rubber to make sealed strip 130, in other embodiments, can also adopt other elaxtic seals such as latex to make sealed strip 130, be not specifically limited herein.
The general glass or polycarbonate (PC) or polymethylmethacrylate (PMMA) material of adopting of upper plate 110 is made, and can also adopt other material in other embodiments, as long as this material has good light transmission, is not specifically limited herein.The general transparent material that adopts of lower plate 120 is made, glass for example, but be also not limited in other embodiments transparent material, as long as can complete its corresponding function, be not specifically limited herein.
Please refer to Fig. 3, on this in-situ polymerization enzyme chain reaction plate 100, be also provided with deflector.The opening direction upper and lower plates 120 of the cavity 140 that upper plate 110 and lower plate 120 are connected to form is longer than upper plate 110, form deflector, reach and when filling in-situ polymerization enzyme chain reaction reagent 150, facilitate drainage and accurately to the effect of the in-situ polymerization enzyme chain reaction reagent 150 of annotating in cavity 140.In other embodiments, this deflector can also be set to other form, for example when filling in-situ polymerization enzyme chain reaction reagent 150, use funnel to form deflector, as long as can reach drainage and accurately to the effect of the in-situ polymerization enzyme chain reaction reagent 150 of annotating in cavity 140, be not specifically limited herein.
Please refer to Fig. 4, before experiment, need to carry out pre-treatment work.In lower plate 120, on the surface relative with upper plate 110, carry out pre-treatment work, tested cell and sample are placed in and on these lower plate 120 surfaces, carry out sticky, the crosslinked and meshization of protein adhesive and process, the final separation rete 121 that forms the net distribution with certain mesh diameter.When manually annotating in-situ polymerization enzyme chain reaction reagent 150, make complete this separation rete 121 of submergence of in-situ polymerization enzyme chain reaction reagent 150, tested nucleus material is fully contacted with in-situ polymerization enzyme chain reaction reagent 150, be beneficial to the carrying out of in-situ polymerization enzyme chain reaction.
On this in-situ polymerization enzyme chain reaction plate 100, be also provided with the sealing cover 141 for sealed cavity 140.Sealing lid 141 can be formed by cover plate, can be also one deck flexible sealing lid.This flexible sealing lid is at one deck oil film then forming to the lightweight oil of annotating in cavity 140 again after the complete in-situ polymerization enzyme chain reaction of the interior filling of cavity 140 reagent 150.Due to oily light specific gravity, so lightweight oil is in fact the top that swims in in-situ polymerization enzyme chain reaction reagent 150, thereby in-situ polymerization enzyme chain reaction reagent 150 is sealed completely, guaranteed the validity sealing in in-situ polymerization enzyme chain reaction process.And due to the sealing function of sealing cover 141, when experiment, in-situ polymerization enzyme chain reaction reagent 150 can not evaporate, and reacts completely, and experiment volume increase thing can not pollute environment, has promoted the success ratio of experiment.In other embodiments, can also form sealing cover 141 with other fluent materials, such as whiteruss etc., but density that need to this fluent material is less than the density of water, is not specifically limited herein.When the two ends of sealed strip 130 extend to the edge of upper plate 110, because lower plate 110 is longer than upper plate 120 on this opening direction, so can be directly one deck glass cover-plate formation sealing cover 141 be set in the edge of upper plate 120; When the two ends of sealed strip 130 do not extend to the edge of upper plate 110, one deck glass cover-plate can be set on the two ends of sealed strip 130 and form sealing cover 141.In other embodiments, can also use other cover plates to form sealing cover 141.When using cover plate to form sealing cover 141, the position that sealing lid 141 arranges need be determined according to the position at the two ends of sealed strip 130.
Please refer to Fig. 3, when using this in-situ polymerization enzyme chain reaction plate 100 to test, place it on worktable, in-situ polymerization enzyme chain reaction plate 100 and the angled α of sea line X-shaped, so that issuable bubble only appears at upside in experimentation.The formation of this angle [alpha] can be to use the supporter with angle [alpha], can be also that in-situ polymerization enzyme chain reaction plate 100 forms self, can utilize the original position gene-amplificative instrament special module with angle [alpha] to form.This angle [alpha] can be 45 ° or 90 °, can be also that non-level other are arbitrarily angled.
Above-mentioned in-situ polymerization enzyme chain reaction plate 100 has a cavity 140 that holds in-situ polymerization enzyme chain reaction reagent 150, at the interior filling in-situ polymerization of this cavity 140 enzyme chain reaction reagent 150 until rete 121 is separated in submergence, then the lightweight oil of then annotating forms sealing cover 141 and swims in in-situ polymerization enzyme chain reaction reagent 150 tops, thereby in-situ polymerization enzyme chain reaction reagent is sealed completely.It is simple to operate when such in-situ polymerization enzyme chain reaction plate 100 is used, and its good stopping property has guaranteed in-situ polymerization enzyme chain reaction reagent 150 sufficient reactings, thereby make experiment there is good amplification, experiment amplified production can not evaporate, not pollute the environment, thereby promoted the success ratio of experiment, the diagnosis of specific gene and pathogenic agent is guaranteed, makes in-situ polymerization enzyme chain reaction technology be able to reliability application and popularization.
The above embodiment has only expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but can not therefore be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (10)
1. an in-situ polymerization enzyme chain reaction plate, it is characterized in that, comprise upper plate and lower plate, surface of described upper plate is provided with sealed strip, described upper plate and described lower plate by described sealed strip be connected to form have opening for holding the cavity of pcr reagent, described in-situ polymerization enzyme chain reaction plate also comprises sealing the sealing cover of described cavity.
2. in-situ polymerization enzyme chain reaction plate according to claim 1, is characterized in that, the surface relative with described upper plate of described lower plate is provided with the separation rete with net distribution that adopts bonding technique and protein cross technology to form.
3. in-situ polymerization enzyme chain reaction plate according to claim 1, is characterized in that, described upper plate adopts transparent material to make.
4. in-situ polymerization enzyme chain reaction plate according to claim 1, is characterized in that, the fluent material that described sealing cover is less than water by density forms.
5. in-situ polymerization enzyme chain reaction plate according to claim 4, is characterized in that, described fluent material is oil or whiteruss.
6. in-situ polymerization enzyme chain reaction plate according to claim 1, is characterized in that, described sealed strip is made by elaxtic seal.
7. in-situ polymerization enzyme chain reaction plate according to claim 1, is characterized in that, described in-situ polymerization enzyme chain reaction plate is placed while using and sea line formation angle, described angle at 0 degree between 180 degree.
8. in-situ polymerization enzyme chain reaction plate according to claim 1, is characterized in that, described lower plate is made by glass or polycarbonate or polymethylmethacrylate.
9. in-situ polymerization enzyme chain reaction plate according to claim 1, is characterized in that, on described in-situ polymerization enzyme chain reaction plate, also has deflector, and described deflector is to be extended and form on the opening direction of described cavity by described lower plate.
10. according to the in-situ polymerization enzyme chain reaction plate described in any one in claim 1～9, it is characterized in that, that described sealed strip is while connecting upper plate and lower plate is U-shaped, V-type or C type.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310674274.4A CN103642674A (en) | 2013-12-11 | 2013-12-11 | In situ polymerase chain reaction plate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310674274.4A CN103642674A (en) | 2013-12-11 | 2013-12-11 | In situ polymerase chain reaction plate |
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| CN103642674A true CN103642674A (en) | 2014-03-19 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201310674274.4A Pending CN103642674A (en) | 2013-12-11 | 2013-12-11 | In situ polymerase chain reaction plate |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105400693A (en) * | 2015-12-15 | 2016-03-16 | 上海海洋大学 | Flat plate isothermal nucleic acid amplification chip |
| CN105400692A (en) * | 2015-12-15 | 2016-03-16 | 上海海洋大学 | Isothermal nucleic acid amplification device and isothermal nucleic acid amplification experimental method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4985206A (en) * | 1987-09-30 | 1991-01-15 | Shandon Scientific Limited | Tissue processing apparatus |
| CN1095759A (en) * | 1993-02-16 | 1994-11-30 | 帕金-埃尔默有限公司 | The in-situ polymerization PCR amplification system |
| US5436129A (en) * | 1989-11-17 | 1995-07-25 | Gene Tec Corp. | Process for specimen handling for analysis of nucleic acids |
| CN203613182U (en) * | 2013-12-11 | 2014-05-28 | 苏州东胜兴业科学仪器有限公司 | In-situ polymerase chain reaction plate |
-
2013
- 2013-12-11 CN CN201310674274.4A patent/CN103642674A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4985206A (en) * | 1987-09-30 | 1991-01-15 | Shandon Scientific Limited | Tissue processing apparatus |
| US5436129A (en) * | 1989-11-17 | 1995-07-25 | Gene Tec Corp. | Process for specimen handling for analysis of nucleic acids |
| CN1095759A (en) * | 1993-02-16 | 1994-11-30 | 帕金-埃尔默有限公司 | The in-situ polymerization PCR amplification system |
| CN203613182U (en) * | 2013-12-11 | 2014-05-28 | 苏州东胜兴业科学仪器有限公司 | In-situ polymerase chain reaction plate |
Non-Patent Citations (1)
| Title |
|---|
| 衷平海: "《实用塑料制品生产技术及应用配方》", 31 January 2005, article "密封条类塑料制品", pages: 379-382 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105400693A (en) * | 2015-12-15 | 2016-03-16 | 上海海洋大学 | Flat plate isothermal nucleic acid amplification chip |
| CN105400692A (en) * | 2015-12-15 | 2016-03-16 | 上海海洋大学 | Isothermal nucleic acid amplification device and isothermal nucleic acid amplification experimental method |
| CN105400692B (en) * | 2015-12-15 | 2017-06-27 | 上海海洋大学 | Isothermal nucleic acid amplification device and isothermal nucleic acid amplification experimental technique |
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Application publication date: 20140319 |