CN1766581A - Safety nucleic acid electrophoresis fluorimeter - Google Patents
Safety nucleic acid electrophoresis fluorimeter Download PDFInfo
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- CN1766581A CN1766581A CN 200510086789 CN200510086789A CN1766581A CN 1766581 A CN1766581 A CN 1766581A CN 200510086789 CN200510086789 CN 200510086789 CN 200510086789 A CN200510086789 A CN 200510086789A CN 1766581 A CN1766581 A CN 1766581A
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- nucleic acid
- electrophoresis
- light source
- gel electrophoresis
- source chamber
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Abstract
The invention relates to a nucleotide separating and testing technology, which is characterized in that the phosphorescence device comprises: a gel electrophoresis tank and a light source room, wherein the exciting shine skin and the average-shine skin inside the light source room are separately fixed on the upper and lower part of the electrophoresis tank bottom plate middle part port; the sample bed plate is fixed on the exciting shine skin; the phosphorescence observing plate is on the top of each electrophoresis tank wall; the electrode bed plates are fixed on the left and right side of the electrophoresis tank bottom plate; the two electrode wires are connected with the electrode power. The white light of the light source room through the laser shine skin generates the exciting light with the wavelength 450nm-510nm. The phosphorescence dye combines to the nucleotide sends the phosphorescence with wavelength 520nm-580nm which can real time observes the location of the nucleotide migration after the filtering of phosphorescence observing sheet.
Description
Technical field
Generally, the present invention relates to the separation detection of nucleic acid.More particularly, the present invention relates to a kind of like this system, during it separates nucleic acid by gel electrophoresis and detects, use the non-toxic, environmental friendly fluorescent dye, thereby make the testing process safety and environmental protection that becomes, the sensitivity of detection is improved.More particularly, the invention still further relates to a kind of like this system, the optical excitation dyestuff of the wavelength 450～510nm of its generation and the compound of nucleic acid produce the fluorescence of 520～580nm, the swimming position of unlabeled nucleic acid, and the light of the various wavelength that relate in the observation process is safe and harmless to human body.
Background technology
Ethidium bromide (EB) is the conventional dyes that separate nucleic acid detects, and nucleic acid samples combines with EB and behind agarose gel electrophoresis, can be by the Fluirescence observation that ultraviolet excitation nucleic acid-the EB compound sends and the size or the purity of definite kernel acid fragment.Because the pyridine of bromine second is combined on the base-pair of DNA chain, thereby has very high toxicity and carcinogenicity, and human body and surrounding environment are all had very big harm.Ultraviolet light is to observe the nucleic acid electrophoresis result, cuts the essential means that glue reclaims nucleic acid fragment, the very easily eyes of harm people and health in the operation.The laboratory concentrates on detection of nucleic acids the platform or the zone of a danger usually, causes greatly inconvenience of research work.
A major defect of existing detection of nucleic acids instrument or system is that the dyestuff EB that is used to detect nucleic acid has high toxicity and carcinogenicity, and operator's operating process and waste disposal all had strict demand, and not high to the detection sensitivity of nucleic acid.
Another major defect of existing instrument is the gel behind the electrophoresis must be observed nucleic acid on ultraviolet transilluminator a separating effect, and ultraviolet ray has very big harm to human body skin and eyes.
Another significant drawbacks of existing system is the testing process complexity, and gel takes out to place on the UV transilluminator and detects behind the electrophoresis tank electrophoresis, can not accomplish real-time detection.
The gel electrophoresis of nucleic acid is the requisite research means of life science.One of them basic work is to separate and the interested nucleic acid of purifying, and in the process of separate nucleic acid purifying, quick, convenient, high sensitivity, nontoxic, free of contamination detection of nucleic acids instrument or device seem particularly important.
Summary of the invention
A fundamental purpose of the present invention provides a kind of nucleic acid gel electrophoresis apparatus, and the nucleic acid dye of its use is nontoxic to human body, environmental sound.
Another object of the present invention provides a kind of nucleic acid detected through gel electrophoresis device, and its exciting light that adopts is the blue light of wavelength 450～510nm, and the wavelength of fluorescence that inspires is 520～580nm, and is harmless.
Another object of the present invention provides a kind of nucleic acid detected through gel electrophoresis device, and its structure comprises the parts of gel electrophoresis groove, generation exciting light, the parts of observation electrophoresis result, realizes real-time monitored, simplifies testing process.
Principle of the present invention is such, all tabula rasa becomes the white light of relatively concentrating that electricity-saving lamp sends into the white face light that is evenly distributed, and this white face light has only the transmittance of 450～510nm wavelength to come out after exciting tabula rasa, the light of this wavelength coverage is exciting light, and the light of other wavelength is all filtered.Produce the fluorescence of 520～580nm behind the optical excitation nucleic acid-fluorescent dye of 450～510nm wavelength, the swimming position of nucleic acid in gel can detect by an unaided eye after with the optical filter of transmission peak wavelength 520～580nm exciting light being filtered.The present invention does not produce deleterious effect to operating personnel and environment, and greatly improves the sensitivity of detection of nucleic acids, to the human zero damage owing to adopt non-toxic dye.The present invention makes one with electrophoresis tank and fluoroscopic examination, has simplified operating process, has realized real-time detection.The light of 450～510nm and 520～580nm is harmless.
The invention is characterized in: this luminoscope contains: the gel electrophoresis groove, and there is opening at the middle part of this trough floor, and the ring limit is arranged on this opening, and each limit on this ring limit all is parallel to each wall of this electrophoresis tank; Excite tabula rasa, be installed on the ring limit of described gel electrophoresis trough floor opening part, this wavelength coverage that excites the transmitted light of tabula rasa is 450nm～510nm;
Sample splint, the limited block by two ends are installed on described exciting on the tabula rasa on this sample splint, and the compound of nucleic acid and non-toxic, environmental friendly fluorescent dye is arranged;
The Fluirescence observation plate is " recessed " font, the anti-lid on described electrophoresis tank, and bonding the pressing plate of " recessed " font in this Fluirescence observation plate center concave surface, the abutting edge is the ring limit of this pressing plate; The wavelength coverage of this Fluirescence observation plate transmitted light is 520nm～580nm, i.e. fluorescence;
Light source chamber, be positioned at the below of described gel electrophoresis trough floor, this light source chamber and described gel electrophoresis groove are made one, the top of this light source chamber is the base plate of described gel electrophoresis groove, and this light source chamber is launched the white face light that is evenly distributed by the opening part of described gel electrophoresis trough floor to the described tabula rasa that excites;
The pair of electrodes supporting plate, the electrode wires on this supporting plate links to each other with electrode supply in the described light source chamber; Described pair of electrodes supporting plate lays respectively at the base plate and arranged on left and right sides of described gel electrophoresis groove.
Basic, apparatus of the present invention are designed to after compound to non-toxic dye and nucleic acid carries out gel electrophoresis, directly observe the fluorescence of wavelength 520～580nm by the observation parts, thereby with electrophoresis with observe two links unite two into one, have realized real-time monitored.The wavelength of observation parts transmitted light is 520～580nm.
Use proof: the present invention has environmental protection, safety, finishes the advantage of the separation and the observation of nucleic acid efficiently, simply and in real time.
Description of drawings:
Fig. 1 is the structural profile stereographic map of the instrument according to the present invention.
Fig. 2 is the rear end top perspective view of Fig. 1 of the instrument according to the present invention.
Fig. 3 is an another kind of version of the present invention, has represented that electrophoresis tank and light source chamber separate the structure of manufacturing and the assembly relation of electrophoresis tank and light source chamber.
Fig. 4 and Fig. 5 are the electrophoresis tank structural perspective.
Fig. 6 is electrode tip holder and electrode wires structure and stereographic map is installed.
Fig. 7 is a sample splint triangle limited block structural perspective.This figure has intercepted a sample splint part, and the intercepting direction is with accompanying drawing 1.
Fig. 8 is equal tabula rasa and light source chamber top board bonded structure synoptic diagram.
Fig. 9 is electricity-saving lamp and circuit and light source chamber base plate mounting structure synoptic diagram.
Figure 10 observes the covering plate structure synoptic diagram.
Figure 11 is the pressure plate structure synoptic diagram of observing in the cover plate.
Embodiment
The object of the present invention is achieved like this, and a kind of non-toxic, environmental friendly real-time monitored nucleic acid fluorescent electrophoresis apparatus comprises electrophoresis tank, light source chamber and observation cover plate.Electrophoresis tank is positioned at the top of light source chamber, and the base plate of electrophoresis tank is the top board of light source chamber, observes cover plate and is positioned at the electrophoresis tank top.Electrophoresis tank comprises electrophoresis tank bottom, excite tabula rasa, electrode, sample splint, the electrophoresis tank bottom middle part is for exciting tabula rasa, and sample splint is placed on and excites on the tabula rasa, bottom land one opposite side has the pair of electrodes seat, two electrode wires are installed in parallel in the groove of electrode tip holder, and an end of electrode wires passes electrophoresis tank bottom and enters light source chamber, link with electrophoresis power socket and electrophoresis switch; Light source chamber comprises the light source chamber top board, equal tabula rasa, electricity-saving lamp and circuit, saving lamp power source socket, electrophoresis power socket, electricity-saving lamp electrophoresis protective tube.Observe cover plate and comprise Fluirescence observation plate and pressing plate, pressing plate is positioned at below, Fluirescence observation plate middle part.
Electrophoresis tank is made by lighttight ordinary hot thermoplastic plastic, thermosetting plastics, engineering plastics or organic glass.Open a rectangular opening at electrophoresis tank bottom middle part, the ring limit of this opening with excite tabula rasa bonding, constitute the electrophoresis tank bottom of sealing.A pair of diameter number millimeter is arranged at an opposite side place of base plate, and high several millimeters small column is used for bonding electrode tip holder.An aperture parallel with cylinder axis is arranged on small column, and the plate of directly revealing the exact details feeds light source chamber, is used for electrode wires is fed light source chamber.Have several grooves at the top of electrophoresis tank wall, be used for discharging the steam that produces in the electrophoresis process.
Exciting tabula rasa is to be made by organic glass or optical filter or thermoplastics, and thickness number millimeter is shaped as rectangle, and transmitted light wavelength 450～510nm sticks with glue with the ring limit at electrophoresis tank bottom middle part and to connect.
Light source chamber is made by lighttight ordinary hot thermoplastic plastic, thermosetting plastics, engineering plastics or organic glass, can be made into integral body with electrophoresis tank, also can make respectively, connects with screw.Open a rectangular opening at light source chamber top board middle part, the ring limit of this opening is with all tabula rasa is bonding.
All tabula rasa is to be made by organic glass or optical filter, and transmitted light is a white light, and thickness number millimeter is shaped as rectangle, size with excite tabula rasa identical, be positioned at and excite under the tabula rasa.
Observe cover plate by the Fluirescence observation plate with pressing plate is bonding forms.The Fluirescence observation plate is made by organic glass or optical filter, can filter the light of 450～510nm.Pressing plate is made concave character type by the transparent whites organic glass, and its concave character type four limits are bonded in Fluirescence observation plate middle part.During use, cover at the electrophoresis tank top with four limits of Fluirescence observation plate concave character type, pressing plate then immerses in the electrophoresis liquid, and pressing plate is positioned at and excites directly over tabula rasa and the sample splint.Can pass through pressing plate observation sample electrophoresis situation, can prevent that again the steam that electrophoresis produces from condensing on the Fluirescence observation plate.
With reference to the accompanying drawings 1 and Fig. 2, instrument of the present invention preferably is rectangular parallelepiped generally, include electrophoresis tank 1, light source chamber 2, observe cover plate 3, panel 4, sample splint 8, and preferably with electrophoresis tank and light source chamber from making one in appearance.This observation of use instrument panel 3 covers the wall top at electrophoresis tank 1, is sample splint 8 below observing panel 3, light source chamber 2 be positioned at electrophoresis tank 1 below.
With reference to the accompanying drawings 3, another kind of preferred structure of the present invention is that electrophoresis tank is to separate with light source chamber, and the two relies on flange 19 and one group of hole 20 usefulness to be threaded.
Electrophoresis tank 1 includes and excites tabula rasa 5, cell body 6, electrode wires and electrode wires supporting plate 7.
With reference to the accompanying drawings 4 and Fig. 5, cell body 6 includes at the base plate middle part an oblong openings, ring limit 29 is arranged on this opening, be used for installing and excite tabula rasa 5, cell body bottom surface corner has the cylinder 22 of a pair of installing electrodes supporting plate 7, the one group of logical vapour groove 28 in cell body wall top, the top standing groove 32 of the long a pair of cell wall of cell body.The limited block 24 of electrode supporting plate 7 is placed in the stopper slot 23 of cell body base plate.Cell body 6 is preferably made with the thermoplastic plastic injecting method, also can use the engineering plastics manufacturing, or the organic glass manufacturing.Cylinder 22 also can be quadrilateral or triangle cylinder.Ring limit 29 is wide several millimeters, exceeds cell body bottom surface 5～15mm.The limit wall of a pair of cylinder 22 has the hole 21 of 0.4～1mm size, and this hole is directly led in the light source chamber 2.
With reference to the accompanying drawings 6, the structure of electrode supporting plate 7 comprises installing ring 25, limited block 24 and electrode wires mounting groove 26.The stretching electrode wires mounting groove 26 of putting into of electrode wires, the electrode wires other end passes hole 21, enters in the light source chamber 2.Electrode supporting plate 7 is connected on the cylinder 22 by installing ring 25 usefulness gum covers, and is pressed on the bottom surface of cell body 6, and limited block 24 places stopper slot 23, to keep supporting plate 7 parallel with cell body wall 31.
With reference to the accompanying drawings 2, panel 4 includes the switch 15 of electricity-saving lamp 11, the switch 16 of electrode 7.Two switches are water proof switch.
With reference to the accompanying drawings 1, Fig. 8 and Fig. 9, light source chamber 2 comprises light source chamber 9, all tabula rasa 10, base plate 14, electricity-saving lamp and circuit 11, saving lamp power source protective tube 12, electrode supply socket 13.
Light source chamber 9 is preferably made one with cell body 6, also can separately make by accompanying drawing 3.The top board of light source chamber 9 is the base plate of cell body 6.There is an oblong openings at top board middle part in light source chamber 9, and ring limit 37 is arranged on this opening, is positioned under the ring limit 29, is used for installing equal tabula rasa 10.
All tabula rasa 10 transmitted lights are white light, and by organic glass or glass filter manufacturing, thickness 1～5mm is rectangle.All tabula rasa 10 sticks with glue on the ring limit 37 that is connected on light source chamber 9.
The end that two electrode wires penetrate light source chamber is connected on the electrode supply socket 13.
With reference to the accompanying drawings 7, sample splint 8 respectively has two triangle limited blocks 27 by white transparent organic glass or white transparent thermoplastics manufacturing at two end faces, and the standing groove 32 of limited block 27 on cell body 6 limits the play of sample splint 8 at in-plane.
With reference to the accompanying drawings 10 and Figure 11, observe cover plate 3 and comprise Fluirescence observation plate 17 and pressing plate 18.Fluirescence observation plate 17 usefulness transparent organic glass or thermoplastics are made concave character type, and its transmitted light wavelength is 520～580nm, covers on electrophoresis tank 6.White transparent organic glass of pressing plate 18 usefulness or white transparent thermoplastics are made concave character type.The ring limit 36 of pressing plate 18 bonds together with the concave surface of Fluirescence observation plate 17, and the position is at the positive middle part of Fluirescence observation plate 18.
During use, sample splint 8 is placed electrophoresis tank 1 together with the gel on it, cover and observe cover plate 3, plug the power lead and the electrophoresis power line of electricity-saving lamp 11, press the electrode switch 16 on the lower panel 4, begin sample electrophoresis, electrophoresis has been pressed switch 16 after finishing, and closes electrophoresis, press down switch 15 then, can be from observing the top Direct observation or the sample amplifying nucleic acid of taking pictures of cover plate 3.Operating process is simple, safety, efficient, environmentally safe, and the operator need not to do any security protection.Thereby according to the present invention, provide a kind of instrument, can be safely, efficient, environmental protection, finish the electrophoresis and the observation process of nucleic acid simply, in real time, satisfy foregoing purpose.Be appreciated that all terms used herein just can not limit the present invention in order to explain.Though foregoing specific embodiments explanation the present invention, according to top described, those skilled in the art can clearly dream up many interchangeable embodiments, distortion and improvement.So the present invention includes all the scope of the invention and the design in and below claim in replaceable scheme, embodiment, distortion and improvement.
Claims (11)
1, safety nucleic acid electrophoresis fluorimeter is characterized in that, this luminoscope contains: the gel electrophoresis groove, and there is opening at the middle part of this trough floor, and the ring limit is arranged on this opening, and each limit on this ring limit all is parallel to each wall of this electrophoresis tank; Excite tabula rasa, be installed on the ring limit of described gel electrophoresis trough floor opening part, this wavelength coverage that excites the transmitted light of tabula rasa is 450nm～510nm;
Sample splint, the limited block by two ends are installed on described exciting on the tabula rasa on this sample splint, and the compound of nucleic acid and non-toxic, environmental friendly fluorescent dye is arranged;
The Fluirescence observation plate is " recessed " font, the anti-lid on described electrophoresis tank, and bonding the pressing plate of " recessed " font in this Fluirescence observation plate center concave surface, the abutting edge is the ring limit of this pressing plate; The wavelength coverage of this Fluirescence observation plate transmitted light is 520nm～580nm, i.e. fluorescence;
Light source chamber, be positioned at the below of described gel electrophoresis trough floor, this light source chamber and described gel electrophoresis groove are made one, the top of this light source chamber is the base plate of described gel electrophoresis groove, and this light source chamber is launched the white face light that is evenly distributed by the opening part of described gel electrophoresis trough floor to the described tabula rasa that excites;
The pair of electrodes supporting plate, the electrode wires on this supporting plate links to each other with electrode supply in the described light source chamber; Described pair of electrodes supporting plate lays respectively at the base plate and arranged on left and right sides of described gel electrophoresis groove.
2, safety nucleic acid electrophoresis fluorimeter according to claim 1, it is characterized in that, described electrode supporting plate comprises installing ring, the limited block that is positioned at the end and is positioned at the mounting groove of middle electrode wires, this installing ring is connected on the cylinder with gum cover, enters described light source chamber and this cylinder is arranged in the electrode wires of this mounting groove on the base plate of described gel electrophoresis groove by the line hole of opening on this cylinder; This limited block place one parallel with described gel electrophoresis groove cell body side plate wall and open stopper slot on described gel electrophoresis trough floor.
3, safety nucleic acid electrophoresis fluorimeter according to claim 1, it is characterized in that, described light source chamber contains: light source chamber, all tabula rasa, light source chamber base plate, electricity-saving lamp and circuit, saving lamp power source protective tube and electrode supply socket, this equal tabula rasa are positioned at and describedly excite under the tabula rasa and be bonded on the bottom surface, ring limit of described gel electrophoresis trough floor opening part.
4, safety nucleic acid electrophoresis fluorimeter according to claim 1 is characterized in that, described light source chamber and gel electrophoresis groove are discrete, and the centre connects by web member with flange.
5, safety nucleic acid electrophoresis fluorimeter according to claim 1 is characterized in that, described gel electrophoresis groove is cuboid, with thermoplastics or engineering plastics or organic glass manufacturing.
6, safety nucleic acid electrophoresis fluorimeter according to claim 1 is characterized in that, the described tabula rasa that excites is rectangle, with transparent organic glass or thermoplastics or optical filter manufacturing.
7, safety nucleic acid electrophoresis fluorimeter according to claim 1 is characterized in that, described Fluirescence observation plate is made with transparent organic glass or glass filter or thermoplastics.
8, safety nucleic acid electrophoresis fluorimeter according to claim 1 is characterized in that, described pressing plate is made with white transparent organic glass or thermoplastics or glass.
9, safety nucleic acid electrophoresis fluorimeter according to claim 3 is characterized in that, described equal tabula rasa is rectangle, and makes with milky white formula organic glass or opal glass optical filter, so that white face light to be provided.
10, safety nucleic acid electrophoresis fluorimeter according to claim 1 is characterized in that, there is one group of air channel on the wall top of described gel electrophoresis groove cell body, the steam that produces in order to discharge heated by electrodes electricity solution.
11, safety nucleic acid electrophoresis fluorimeter according to claim 1 is characterized in that, on the long a pair of cell wall of described gel electrophoresis groove the standing groove of placing described Fluirescence observation plate is arranged.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100867898A CN100473975C (en) | 2005-11-04 | 2005-11-04 | Safety nucleic acid electrophoresis fluorimeter |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100867898A CN100473975C (en) | 2005-11-04 | 2005-11-04 | Safety nucleic acid electrophoresis fluorimeter |
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| Publication Number | Publication Date |
|---|---|
| CN1766581A true CN1766581A (en) | 2006-05-03 |
| CN100473975C CN100473975C (en) | 2009-04-01 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100867898A Expired - Fee Related CN100473975C (en) | 2005-11-04 | 2005-11-04 | Safety nucleic acid electrophoresis fluorimeter |
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| CN (1) | CN100473975C (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100458408C (en) * | 2007-01-18 | 2009-02-04 | 深圳大学 | Polaroid type nucleic acid electrophoresis apparatus |
| CN101949886A (en) * | 2010-08-27 | 2011-01-19 | 亚亚科技股份有限公司 | Real-time fluorescence electrophoresis apparatus |
| CN103245640A (en) * | 2012-02-08 | 2013-08-14 | 鸿林堂科技股份有限公司 | Double-source lamp box experiment system |
| CN108535221A (en) * | 2017-03-03 | 2018-09-14 | 广州博鹭腾仪器仪表有限公司 | A kind of new type gel imaging system |
-
2005
- 2005-11-04 CN CNB2005100867898A patent/CN100473975C/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100458408C (en) * | 2007-01-18 | 2009-02-04 | 深圳大学 | Polaroid type nucleic acid electrophoresis apparatus |
| CN101949886A (en) * | 2010-08-27 | 2011-01-19 | 亚亚科技股份有限公司 | Real-time fluorescence electrophoresis apparatus |
| CN103245640A (en) * | 2012-02-08 | 2013-08-14 | 鸿林堂科技股份有限公司 | Double-source lamp box experiment system |
| CN103245640B (en) * | 2012-02-08 | 2015-09-30 | 鸿林堂科技股份有限公司 | Two luminous source lamp box experimental system |
| CN108535221A (en) * | 2017-03-03 | 2018-09-14 | 广州博鹭腾仪器仪表有限公司 | A kind of new type gel imaging system |
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| Publication number | Publication date |
|---|---|
| CN100473975C (en) | 2009-04-01 |
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Granted publication date: 20090401 Termination date: 20091204 |