CN110095614A - A kind of method of quick detection genetically modified plants transgenic protein B t-Cry1AbAc - Google Patents
A kind of method of quick detection genetically modified plants transgenic protein B t-Cry1AbAc Download PDFInfo
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- CN110095614A CN110095614A CN201910445544.1A CN201910445544A CN110095614A CN 110095614 A CN110095614 A CN 110095614A CN 201910445544 A CN201910445544 A CN 201910445544A CN 110095614 A CN110095614 A CN 110095614A
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- stirring rod
- cry1abac
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- 238000000034 method Methods 0.000 title claims abstract description 28
- 238000001514 detection method Methods 0.000 title claims abstract description 20
- 108010042653 IgA receptor Proteins 0.000 title claims abstract description 16
- 102100034014 Prolyl 3-hydroxylase 3 Human genes 0.000 title claims abstract description 16
- 230000009261 transgenic effect Effects 0.000 title claims abstract description 16
- 238000012360 testing method Methods 0.000 claims abstract description 79
- 238000003756 stirring Methods 0.000 claims abstract description 36
- 239000000243 solution Substances 0.000 claims abstract description 33
- 238000001035 drying Methods 0.000 claims abstract description 28
- 238000004140 cleaning Methods 0.000 claims abstract description 24
- 239000007864 aqueous solution Substances 0.000 claims abstract description 18
- 238000003780 insertion Methods 0.000 claims abstract description 8
- 230000037431 insertion Effects 0.000 claims abstract description 8
- 239000007787 solid Substances 0.000 claims abstract description 7
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- 239000008187 granular material Substances 0.000 claims description 9
- 239000008367 deionised water Substances 0.000 claims description 8
- 229910021641 deionized water Inorganic materials 0.000 claims description 8
- 230000000249 desinfective effect Effects 0.000 claims description 8
- 238000004090 dissolution Methods 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 8
- 241000973497 Siphonognathus argyrophanes Species 0.000 claims description 4
- 230000008676 import Effects 0.000 claims description 4
- 239000002245 particle Substances 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 3
- 230000002000 scavenging effect Effects 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 description 42
- 108090000623 proteins and genes Proteins 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 8
- 230000001376 precipitating effect Effects 0.000 description 4
- 238000002331 protein detection Methods 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000011435 rock Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 108700019146 Transgenes Proteins 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
Abstract
The invention discloses the methods of quickly detection genetically modified plants transgenic protein B t-Cry1AbAc a kind of, include the following steps: that cleaning, drying will be carried out to measuring plants, after cleaning, drying, it will pour into grinding device and crush to measuring plants, grinding time 5-10 minutes, container and stirring rod are carried out disinfection, it will be poured into container by what is crushed to measuring plants again, it is slowly uniform that aqueous solution is added, and it is stirred simultaneously with stirring rod, covered container lid, container is rocked into 15-20S up and down, by solution left standstill 5-8 minutes, solid deposits layering, take upper solution in test tube, dismantle test paper, vertically downward by the sample end of test paper, insertion is equipped in the test tube of solution, it impregnates 1-2 minutes, take out test paper, start timing, after 5-10 minutes, Read test result.
Description
Technical field
The present invention relates to gene protein detection technique fields, more particularly to a kind of quickly detection genetically modified plants transgenic
The method of protein B t-Cry1AbAc.
Background technique
The success of genetically modified plants greatly improves the yield of planting, as planting for the plants such as corn and soybean
To popularization.Contain transgene protein in the product that genetically modified plants are produced, genetically modified plants need in product containing turning base
Because albumen is detected, the excellent of genetically modified plants is judged.Existing detection method is complicated for operation, needs using more device
Tool, testing cost are high.
Summary of the invention
The invention proposes the methods of quickly detection genetically modified plants transgenic protein B t-Cry1AbAc a kind of, with solution
Certainly the problems mentioned above in the background art.
The invention proposes the methods of quickly detection genetically modified plants transgenic protein B t-Cry1AbAc a kind of, including
Following steps:
S1: will carry out cleaning, drying to measuring plants, after cleaning, drying, will pour into grinding device and carry out to measuring plants
It crushes, grinding time 5-10 minutes；
S2: container and stirring rod are carried out disinfection, then will be poured into container by what is crushed to measuring plants, slowly uniform
Aqueous solution is added, and is stirred simultaneously with stirring rod；
S3: container is rocked 15-20S up and down, so that grain dissolution is in aqueous solution by covered container lid；
S4: by solution left standstill 2-4 minutes, solid deposits were layered；
S5: taking upper solution in test tube, dismantles test paper, vertically downward by the sample end of test paper, insertion dress
Have in the test tube of solution, impregnate 1-2 minutes, the inserting step of test paper:
A, the direction for adjusting test paper is moved at test tube mouth 1-3cm so that the sample end of test paper is downward；
B, slowly mobile test test paper again, so that the sample end of test paper gradually invades in invisible spectro solution；
C, handheld test test paper remains stationary, so that test paper gos deep into the depth in solution and keeps a certain value range；
S6: taking out test paper, starts timing, after 5-10 minutes, read test result.
Preferably, in the S1, the smashed particle diameter is less than 1 millimeter.
Preferably, in the S1, the cleaning, drying step to measuring plants are as follows:
(1) it will be imported to measuring plants and carry out rotary-cleaning in roller, scavenging period is 3-8 minutes；
(2) after cleaning, it will import in drying box and dry to measuring plants, the temperature in drying box is 50-60 degrees Celsius,
Drying time is 5-10 minutes.
Preferably, in the S2, the aqueous solution is deionized water solution.
Preferably, in the S2, in the stirring rod whipping process, so that the plant granule to be measured in container will not be bonded
Together, plant granule to be measured dissolves in aqueous solution.
Preferably, in the S2, the disinfecting process of the container and stirring rod are as follows:
(1) it will be impregnated 3-5 minutes in container and stirring rod merging hot water, hot water temperature is 95-100 DEG C；
(2) container and stirring rod are taken out, container and stirring rod are placed in ultraviolet lamp and sterilized, disinfecting time 2-5 points
Clock；
(3) container and stirring rod are taken out, is cooled to room temperature.
It preferably, is about 0.3-0.6cm by the depth that test paper sample end is submerged into sample liquid in the S5, it should not
More than the density bullet line of arrow upper end.
The method of quickly detection genetically modified plants transgenic protein B t-Cry1AbAc proposed by the present invention a kind of is beneficial
Effect is:
1, pass through deionized water solution, be free of ion in deionized water solution, reduce the influence detected to gene protein, and same
When be stirred with stirring rod, in stirring rod whipping process so that the plant granule to be measured in container will not bond together, to
Measuring plants grain dissolution is in aqueous solution.
2, vertically downward by the sample end of test paper, insertion is equipped in the test tube of solution, the sample end of paper of having a try by
It gradually invades in invisible spectro solution, the depth for submerging sample liquid is 0.3-0.6cm, ensure that the accuracy of detection.
Specific embodiment
It is next combined with specific embodiments below that the present invention will be further described.
Embodiment 1
The invention proposes the methods of quickly detection genetically modified plants transgenic protein B t-Cry1AbAc a kind of, including
Following steps:
S1: will carry out cleaning, drying to measuring plants, after cleaning, drying, will pour into grinding device and carry out to measuring plants
It crushes, grinding time 5 minutes, cleaning, drying step of the smashed particle diameter less than 1 millimeter, to measuring plants are as follows:
(1) it will be imported to measuring plants and carry out rotary-cleaning in roller, rotary-cleaning can effectively remove the sundries on surface, clearly
Washing the time is 3 minutes；
(2) after cleaning, it will import in drying box and dry to measuring plants, drying the temperature inside the box is 50 degrees Celsius, low temperature
Drying will not destroy the ingredient to measuring plants, and drying time is 5 minutes；
S2: container and stirring rod are carried out disinfection, and avoid bacterial micro-organism from influencing the detection of gene protein, then will pass through powder
Broken pours into container to measuring plants, slowly uniform to be added aqueous solution, and aqueous solution is deionized water solution, in deionized water solution
Without ion, reduce the influence detected to gene protein, and be stirred simultaneously with stirring rod, in stirring rod whipping process, is made
The plant granule to be measured obtained in container will not bond together, and plant granule to be measured dissolves in aqueous solution, container and stirring rod
Disinfecting process are as follows:
(1) it will be impregnated 3 minutes in container and stirring rod merging hot water, hot water temperature is 95 DEG C；
(2) container and stirring rod are taken out, container and stirring rod are placed in ultraviolet lamp and sterilized, disinfecting time 2 minutes；
(3) container and stirring rod are taken out, is cooled to room temperature；
S3: container is rocked 15S up and down, so that grain dissolution is in aqueous solution, rocked up and down uniformly by covered container lid
It rocks, measuring plants grain dissolution, so that the precipitating generated is less；
S4: by solution left standstill 2 minutes, solid deposits layering, solid deposits were being extracted clearly in the bottom end of container
When liquid, there is no precipitatings, so that deposit will not influence gene protein detection；
S5: taking upper solution in test tube, dismantles test paper, vertically downward by the sample end of test paper, insertion dress
Have in the test tube of solution, impregnates 1 minute, be about 0.3cm by the depth that test paper sample end is submerged into sample liquid, do not exceed
The density bullet line of arrow upper end, the inserting step of test paper:
A, the direction for adjusting test paper is moved at test tube mouth 1cm so that the sample end of test paper is downward；
B, slowly mobile test test paper is kept away so that the sample end of test paper gradually invades in invisible spectro solution again
It is excessive to exempt from test paper insertion；
C, handheld test test paper remains stationary, so that test paper gos deep into the depth in solution and keeps a certain value range；
S6: taking out test paper, starts timing, after five minutes, read test result.
Embodiment 2
The invention proposes the methods of quickly detection genetically modified plants transgenic protein B t-Cry1AbAc a kind of, including
Following steps:
S1: will carry out cleaning, drying to measuring plants, after cleaning, drying, will pour into grinding device and carry out to measuring plants
It crushes, grinding time 10 minutes, cleaning, drying step of the smashed particle diameter less than 1 millimeter, to measuring plants are as follows:
(1) it will be imported to measuring plants and carry out rotary-cleaning in roller, rotary-cleaning can effectively remove the sundries on surface, clearly
Washing the time is 8 minutes；
(2) after cleaning, it will import in drying box and dry to measuring plants, drying the temperature inside the box is 60 degrees Celsius, low temperature
Drying will not destroy the ingredient to measuring plants, and drying time is 10 minutes；
S2: container and stirring rod are carried out disinfection, and avoid bacterial micro-organism from influencing the detection of gene protein, then will pass through powder
Broken pours into container to measuring plants, slowly uniform to be added aqueous solution, and aqueous solution is deionized water solution, in deionized water solution
Without ion, reduce the influence detected to gene protein, and be stirred simultaneously with stirring rod, in stirring rod whipping process, is made
The plant granule to be measured obtained in container will not bond together, and plant granule to be measured dissolves in aqueous solution, container and stirring rod
Disinfecting process are as follows:
(1) it will be impregnated 5 minutes in container and stirring rod merging hot water, hot water temperature is 100 DEG C；
(2) container and stirring rod are taken out, container and stirring rod are placed in ultraviolet lamp and sterilized, disinfecting time 5 minutes；
(3) container and stirring rod are taken out, is cooled to room temperature；
S3: container is rocked 20S up and down, so that grain dissolution is in aqueous solution, rocked up and down uniformly by covered container lid
It rocks, measuring plants grain dissolution, so that the precipitating generated is less；
S4: by solution left standstill 4 minutes, solid deposits layering, solid deposits were being extracted clearly in the bottom end of container
When liquid, there is no precipitatings, so that deposit will not influence gene protein detection；
S5: taking upper solution in test tube, dismantles test paper, vertically downward by the sample end of test paper, insertion dress
Have in the test tube of solution, impregnates 2 minutes, be about 0.6cm by the depth that test paper sample end is submerged into sample liquid, do not exceed
The density bullet line of arrow upper end, the inserting step of test paper:
A, the direction for adjusting test paper is moved at test tube mouth 3cm so that the sample end of test paper is downward；
B, slowly mobile test test paper is kept away so that the sample end of test paper gradually invades in invisible spectro solution again
It is excessive to exempt from test paper insertion；
C, handheld test test paper remains stationary, so that test paper gos deep into the depth in solution and keeps a certain value range；
S6: taking out test paper, starts timing, after ten minutes, read test result.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (7)
1. a kind of method of quickly detection genetically modified plants transgenic protein B t-Cry1AbAc, which is characterized in that including following
Step:
S1: will carry out cleaning, drying to measuring plants, after cleaning, drying, will pour into measuring plants and carry out powder in grinding device
It is broken, grinding time 5-10 minutes；
S2: container and stirring rod are carried out disinfection, then will be poured into container by what is crushed to measuring plants, slowly uniform to be added
Aqueous solution, and be stirred simultaneously with stirring rod；
S3: container is rocked 15-20S up and down, so that grain dissolution is in aqueous solution by covered container lid；
S4: by solution left standstill 2-4 minutes, solid deposits were layered；
S5: taking upper solution in test tube, dismantles test paper, and vertically downward by the sample end of test paper, insertion is equipped with molten
In the test tube of liquid, impregnate 1-2 minutes, the inserting step of test paper:
A, the direction for adjusting test paper is moved at test tube mouth 1-3cm so that the sample end of test paper is downward；
B, slowly mobile test test paper again, so that the sample end of test paper gradually invades in invisible spectro solution；
C, handheld test test paper remains stationary, so that test paper gos deep into the depth in solution and keeps a certain value range；
S6: taking out test paper, starts timing, after 5-10 minutes, read test result.
2. the method for quickly detection genetically modified plants transgenic protein B t-Cry1AbAc according to claim 1 a kind of,
It is characterized in that, the smashed particle diameter is less than 1 millimeter in the S1.
3. the method for quickly detection genetically modified plants transgenic protein B t-Cry1AbAc according to claim 1 a kind of,
It is characterized in that, in the S1, the cleaning, drying step to measuring plants are as follows:
(1) it will be imported to measuring plants and carry out rotary-cleaning in roller, scavenging period is 3-8 minutes；
(2) after cleaning, it will import in drying box and dry to measuring plants, the temperature in drying box is 50-60 degrees Celsius, drying
Time is 5-10 minutes.
4. the method for quickly detection genetically modified plants transgenic protein B t-Cry1AbAc according to claim 1 a kind of,
It is characterized in that, the aqueous solution is deionized water solution in the S2.
5. the method for quickly detection genetically modified plants transgenic protein B t-Cry1AbAc according to claim 1 a kind of,
It is characterized in that, in the S2, in the stirring rod whipping process, so that the plant granule to be measured in container will not be bonded in one
It rises, plant granule dissolution to be measured is in aqueous solution.
6. the method for quickly detection genetically modified plants transgenic protein B t-Cry1AbAc according to claim 1 a kind of,
It is characterized in that, in the S2, the disinfecting process of the container and stirring rod are as follows:
(1) it will be impregnated 3-5 minutes in container and stirring rod merging hot water, hot water temperature is 95-100 DEG C；
(2) container and stirring rod are taken out, container and stirring rod are placed in ultraviolet lamp and sterilized, disinfecting time 2-5 minutes；
(3) container and stirring rod are taken out, is cooled to room temperature.
7. the method for quickly detection genetically modified plants transgenic protein B t-Cry1AbAc according to claim 1 a kind of,
It is characterized in that, being about 0.3-0.6cm by the depth that test paper sample end is submerged into sample liquid, not exceeding in the S5
The density bullet line of arrow upper end.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910445544.1A CN110095614A (en) | 2019-05-27 | 2019-05-27 | A kind of method of quick detection genetically modified plants transgenic protein B t-Cry1AbAc |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110564822A (en) * | 2019-09-20 | 2019-12-13 | 中国科学院成都生物研究所 | LAMP technology-based transgenic corn Bt176 related gene detection method and kit |
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