CN202626164U - Device for performing covalent cross-linking on propylene glycol monomethyl acetate (PMA)/ ethyl methacrylate (EMA) and deoxyribonucleic acid (DNA) molecules of dead bacterial cell - Google Patents
Device for performing covalent cross-linking on propylene glycol monomethyl acetate (PMA)/ ethyl methacrylate (EMA) and deoxyribonucleic acid (DNA) molecules of dead bacterial cell Download PDFInfo
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- CN202626164U CN202626164U CN 201220228104 CN201220228104U CN202626164U CN 202626164 U CN202626164 U CN 202626164U CN 201220228104 CN201220228104 CN 201220228104 CN 201220228104 U CN201220228104 U CN 201220228104U CN 202626164 U CN202626164 U CN 202626164U
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- pma
- ema
- covalent cross
- linking
- dna
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Abstract
The utility model discloses a device for performing covalent cross-linking on propylene glycol monomethyl acetate (PMA)/ ethyl methacrylate (EMA) and deoxyribonucleic acid (DNA) molecules of a dead bacterial cell. The device comprises a box body which is provided with a movable door, a halogen lamp switch, an ultraviolet lamp tube switch, and a power master switch; a halogen lamp is arranged on the top in the box body; and at least one ultraviolet lamp tube is arranged on the inner wall of each of two sides of the box body. The device overcomes the defects in the prior art, is convenient to operate, and can perform ultraviolet germicidal irradiation; and light is uniform.
Description
Technical field
The utility model relates to a kind of PMA/EMA of being used for and bacterium dead cell dna molecular covalent cross-linking device.
Background technology
Real time pcr is employed for many years, is used for detecting the pathogenic micro-organism of food, feed or environment.Yet false-positive result appears easily in present PCR detection method, and this has caused the generation of the huge waste and the unnecessary incident of recalling.The reason that this result occurs is that round pcr follows other DNA method the same, all can not separate with dead bacterium what live, has caused round pcr that wrong result is provided.
In order to overcome the weakness of above round pcr, avoid the appearance of false positive results, combine round pcr that sample is handled single nitrine ethidium bromide dyestuff.Single nitrine ethidium bromide combines with dna molecular after getting in the dead bacterial body; Single nitrine ethidium bromide dyestuff makes that bacterium can not be dissolved after dyeing; Opposite is; Dyestuff can not penetrate viable cell, and this makes the testing staff can utilize this improved round pcr to distinguish alive and dead bacterium easily, thereby avoids the appearance of false positive results.
The utility model content
By Given this, the purpose of the utility model provides a kind of PMA/EMA of being used for and bacterium dead cell dna molecular covalent cross-linking device, has overcome the deficiency of prior art, and easy to operate, illumination evenly and can carry out ultraviolet-sterilization.
To achieve these goals, the utility model adopts following technical scheme:
A kind of PMA/EMA and bacterium dead cell dna molecular covalent cross-linking device of being used for; Comprise casing; Wherein, Said casing is provided with dodge gate, halogen lamp switch, ultraviolet lamp tube switch and total power switch, and said casing inner top is provided with halogen lamp, is respectively arranged with at least one ultraviolet lamp tube on the inwall of casing both sides.
Further, described ultraviolet lamp tube is two.
Further, also be provided with visual windows on the said casing.
Further, said halogen lamp is 500 watts.
The beneficial effect of the utility model is:
The utility model can be used for the dna molecular of permanent modification bacterium dead cell, the polymerase chain reaction of blocking dna molecule, thus effectively distinguish the life or death state of bacterium in the sample, reduce the false positive rate of pathogenic bacterium in the PCR test sample.The utility model can provide the high light of about 20000lux; And then it is crosslinked in this device, to carry out illumination after PMA and bacteria suspension are mixed; PMA with the photosensitivity azido group be converted into highly active nitrene radical; And form stable covalency nitrogen carbon back with near the binding site any hydrocarbon reaction, and cause the permanent modification of dead cell dna molecular, block polymerase chain reaction (PCR) amplification of dead cell dna molecular at last.
Other advantages, target and the characteristic of the utility model will be set forth in specification sheets subsequently to a certain extent; And to a certain extent; Based on being conspicuous to those skilled in the art, perhaps can from the practice of the utility model, obtain instruction to investigating of hereinafter.The target of the utility model can realize through the structure that is particularly pointed out in following specification sheets or the accompanying drawing and obtain with other advantages.
Description of drawings
Fig. 1 is the structural representation of the utility model.
Embodiment
As shown in Figure 1; The casing 1 of the utility model is provided with visual windows 3, dodge gate 5, halogen lamp switch 6, ultraviolet lamp tube switch 7 and total power switch 8; Dodge gate 5 is hinged with casing 1; The inwall at casing 1 top is provided with the halogen lamp 4 of 500W, is separately installed with 2, two parallel ultraviolet lamp tubes 2 of two ultraviolet lamp tubes on the inwall of casing 1 both sides and is symmetrical set respectively in the medial surface of casing 1.Halogen lamp 4 provides high light, and ultraviolet lamp tube 2 is used for sterilization, in order to improve sterilization effect, two ultraviolet lamp tubes 2 is installed respectively on the inwall of casing 1 both sides.Observe inner illumination cross-linking effect through visual windows 3.
The utility model can be used for the dna molecular of permanent modification bacterium dead cell, the polymerase chain reaction of blocking dna molecule, thus effectively distinguish the life or death state of bacterium in the sample, reduce the false positive rate of pathogenic bacterium in the PCR test sample.The utility model can provide the high light of about 20000lux; And then it is crosslinked in this device, to carry out illumination after PMA and bacteria suspension are mixed; PMA with the photosensitivity azido group be converted into highly active nitrene radical; And form stable covalency nitrogen carbon back with near the binding site any hydrocarbon reaction, and cause the permanent modification of dead cell dna molecular, block polymerase chain reaction (PCR) amplification of dead cell dna molecular at last.
Use Salmonellas viable cell in the utility model rapid detection feed, adopt following steps:
1) the feed sample is increased bacterium with BPW meat soup, draw the 500ul bacteria suspension in the 1.5mL centrifuge tube, the nitrine bromination of adding 4ul 0.5mg/mL is pyridine solution, and centrifuge tube is placed on the ice chest, and 3min makes public in the covalent cross-linking device;
2) extract DNA of bacteria;
3) pcr amplification Salmonellas invA gene: get premix enzyme and primer mixture in the test kit, the 50ul system is drawn 40ul, adds DNA of bacteria 10 ul that extract and carries out pcr amplification; Amplification condition is: 94 ℃ of preparatory sex change 5min; 94 ℃ of sex change 30sec, 56 ℃ of annealing 30 sec, 72 ℃ are extended 45sec; 33 circulations, last 72 ℃ are extended 10 min eventually;
4) electrophoretic analysis: the PCR product is carried out electrophoresis on 1% sepharose, after electrophoresis finished, gel imaging system carried out imaging analysis.
Explanation is at last; Above embodiment is only unrestricted in order to the technical scheme of explanation the utility model; Other modifications that those of ordinary skills make the technical scheme of the utility model perhaps are equal to replacement; Only otherwise break away from the spirit and the scope of the utility model technical scheme, all should be encompassed in the claim scope of the utility model.
Claims (4)
1. one kind is used for PMA/EMA and bacterium dead cell dna molecular covalent cross-linking device; Comprise casing; It is characterized in that: said casing is provided with dodge gate, halogen lamp switch, ultraviolet lamp tube switch and total power switch; Said casing inner top is provided with halogen lamp, is respectively arranged with at least one ultraviolet lamp tube on the inwall of casing both sides.
2. a kind of PMA/EMA and bacterium dead cell dna molecular covalent cross-linking device of being used for according to claim 1, it is characterized in that: described ultraviolet lamp tube is two.
3. a kind of PMA/EMA and bacterium dead cell dna molecular covalent cross-linking device of being used for according to claim 1 and 2 is characterized in that: also be provided with visual windows on the said casing.
4. a kind of PMA/EMA and bacterium dead cell dna molecular covalent cross-linking device of being used for according to claim 3, it is characterized in that: said halogen lamp is 500 watts.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201220228104 CN202626164U (en) | 2012-05-21 | 2012-05-21 | Device for performing covalent cross-linking on propylene glycol monomethyl acetate (PMA)/ ethyl methacrylate (EMA) and deoxyribonucleic acid (DNA) molecules of dead bacterial cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201220228104 CN202626164U (en) | 2012-05-21 | 2012-05-21 | Device for performing covalent cross-linking on propylene glycol monomethyl acetate (PMA)/ ethyl methacrylate (EMA) and deoxyribonucleic acid (DNA) molecules of dead bacterial cell |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN202626164U true CN202626164U (en) | 2012-12-26 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201220228104 Expired - Fee Related CN202626164U (en) | 2012-05-21 | 2012-05-21 | Device for performing covalent cross-linking on propylene glycol monomethyl acetate (PMA)/ ethyl methacrylate (EMA) and deoxyribonucleic acid (DNA) molecules of dead bacterial cell |
Country Status (1)
| Country | Link |
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| CN (1) | CN202626164U (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103224881A (en) * | 2013-03-27 | 2013-07-31 | 北京博瑞立安生物技术有限公司 | Pre-treatment device and pre-treatment method for viable bacteria detection |
| WO2020136595A1 (en) | 2018-12-25 | 2020-07-02 | Tubitak | Fast and portable microfluidic detection system as an alternative to salmonella's classical culture method |
-
2012
- 2012-05-21 CN CN 201220228104 patent/CN202626164U/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103224881A (en) * | 2013-03-27 | 2013-07-31 | 北京博瑞立安生物技术有限公司 | Pre-treatment device and pre-treatment method for viable bacteria detection |
| CN103224881B (en) * | 2013-03-27 | 2015-01-21 | 北京博瑞立安生物技术有限公司 | Pre-treatment device and pre-treatment method for viable bacteria detection |
| WO2020136595A1 (en) | 2018-12-25 | 2020-07-02 | Tubitak | Fast and portable microfluidic detection system as an alternative to salmonella's classical culture method |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20121226 Termination date: 20140521 |