CN107034140B - A kind of nanogel slant medium and preparation method thereof - Google Patents
A kind of nanogel slant medium and preparation method thereof Download PDFInfo
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- CN107034140B CN107034140B CN201710380512.9A CN201710380512A CN107034140B CN 107034140 B CN107034140 B CN 107034140B CN 201710380512 A CN201710380512 A CN 201710380512A CN 107034140 B CN107034140 B CN 107034140B
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Abstract
A kind of nanogel slant medium of disclosure of the invention and preparation method thereof, belongs to microculture field.With part nanogel particle and the agar powder of Ago-Gel substitution part on the basis of common slant agar culture medium, the agar powder first with nanogel particle substitution 20%-30%, then the agar powder with Ago-Gel substitution 25%-35% are specifically calculated based on mass fraction；The setting time and setting rate of culture medium are effectively delayed, caliper uniformity degree when improving slant medium preparation, also improve the uniform distribution of forces of culture medium nutriment, to which the accuracy of culture and follow-up test result be effectively ensured, avoids being repeated several times and test, shorten test period, reduce error, be conducive to improve the levels of precision of test, in addition, preparation method design of the invention is rationally, it is simple to operate.
Description
Technical field
The present invention relates to microculture fields more particularly to a kind of nanogel slant medium and preparation method thereof.
Background technique
Culture medium is that fermentation process or cell culture of animals are raw for microorganism or animal and plant cell in the process
Nutrient matrix necessary to long, breeding or accumulation metabolite.The ingredient of general culture medium includes: carbon source, nitrogen source, minerals,
And other necessary materials, the substance that such as certain biologies cannot synthesize itself, such as certain amino acid, vitamin or nucleotide.
In addition, being commonly incorporated into pH buffer in culture medium also to maintain stable pH.
It is that the carrier mainly solidified is aided with other nutriments again that solid medium, which is typically all with agar, in the prior art,
But be applied to have certain problems when slant medium, since setting rate of the agar at 40 DEG C -45 DEG C is too fast, solidification
Time is too short to cause when carrying out slant medium preparation, be easy to cause solidification uneven, and difference in thickness is too big up and down on inclined-plane asks
Topic, on the one hand will affect the growing nutrient equilibrium degree of bacterium solution, occurs crawling, too thin ground when on the other hand will cause coating
Occur scratching accuracy problem of the rotten situation to influence later period test when side's scribing line.
Summary of the invention
Goal of the invention: in order to overcome problems of the prior art, the invention proposes a kind of effectively control culture mediums
Setting rate, when improving slant medium preparation caliper uniformity and culture medium nutriment be uniformly distributed problem nanometer it is solidifying
Glue slant medium and preparation method thereof.
Technical solution: in order to solve the above-mentioned technical problem, the technical scheme adopted by the invention is as follows: a kind of nanogel is oblique
Face culture medium uses part nanogel particle and Ago-Gel substitution portion on the basis of common slant agar culture medium
The agar powder divided specifically calculates the agar powder first with nanogel particle substitution 20%-30% based on mass fraction, then uses agar
The agar powder of sugared gel substitution 25%-35%；
The nanogel particle is the preparation of zinc oxide-titanic oxide nano compound material: zinc chloride is dissolved in ten times
In the acid solution of the dehydrated alcohol of volume, in low-temperature plasma reactor react 10-20min after be added titania powder after
Continuous reaction 20-30min, reaction temperature are -12~-6 DEG C, and it is uniform to solution dispersion to take out sufficiently ultrasonic agitation after completion of the reaction；
Adjusting pH is 2-3, after be placed in -5-0 DEG C of standing 12h or more in reaction of low temperature plasma device again, form uniform colloidal sol, further take out
Evaporation drying is changed into gel, obtains nanogel particle.
More preferably, the Ago-Gel is to be formulated with agarose and TAE solution with the mass ratio of 2:3.
More preferably, the nanogel particle is not high in the mass percentage content in addition to cultivating based solvent distilled water
In 50%.
More preferably, the size of the nanogel particle is less than 0.1 μm.
The invention also discloses a kind of above-mentioned preparation methods of nanogel slant medium, include the following steps:
A) the preparation of culture medium base fluid: by the slant agar culture medium formula of required purposes, its in addition to agar powder is weighed
His raw material, dissolution are stirred into culture medium base fluid, adjust and arrive required pH；
B the desired amount of agar calculated) the preparation of culture medium: is added into the culture medium base fluid for having regulated pH
Heating stirring adds nanogel particle to being completely dissolved after powder, Ago-Gel, continues heating stirring to being completely dissolved, presses
Preparation method packing, the inclination solidification of slant agar culture medium obtain purpose culture medium；
C) post-process: heat preservation for standby use after sterilization treatment obtains required nanogel slant medium.
The sterilization treatment concrete mode is that chemical sterilization processing mode living carries out, and the mode of autoclave sterilization can be to agarose
Gel generates destruction.
The utility model has the advantages that a kind of nanogel slant medium provided by the invention and preparation method thereof, belongs to microorganism training
The field of supporting.Part is substituted with part nanogel particle and Ago-Gel on the basis of common slant agar culture medium
Agar powder, specifically calculate the agar powder first with nanogel particle substitution 20%-30% based on mass fraction, then use agarose
The agar powder of gel substitution 25%-35%；The setting time and setting rate for effectively having delayed culture medium improve inclined-plane training
Caliper uniformity degree when supporting base preparation, also improves the uniform distribution of forces of culture medium nutriment, so that training be effectively ensured
Feeding and follow-up test result accuracy is avoided being repeated several times and be tested, shortens test period, reduces error, be conducive to improve
The levels of precision of test, in addition, preparation method design of the invention is rationally, it is simple to operate.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail:
A kind of nanogel slant medium uses part nanogel grain on the basis of common slant agar culture medium
Son and the agar powder of Ago-Gel substitution part, specifically calculate based on mass fraction and are substituted first with nanogel particle
The agar powder of 20%-30%, then the agar powder with Ago-Gel substitution 25%-35%；
Embodiment 1:
A kind of nanogel slant medium specifically includes following ingredient: grape for the culture of staphylococcus aureus
Sugared 20g, peptone 15g, sodium chloride 5g, beef extract 0.5g, agar powder 7g, nanogel particle 6g, Ago-Gel 7g；Add steaming
Distilled water is formulated into 1L；
The nanogel particle is the preparation of zinc oxide-titanic oxide nano compound material: zinc chloride is dissolved in ten times
Titania powder continuation is added after reacting 10min in the acid solution of the dehydrated alcohol of volume, in low-temperature plasma reactor
20min is reacted, reaction temperature is -12 DEG C, and it is uniform to solution dispersion to take out sufficiently ultrasonic agitation after completion of the reaction；Adjusting pH is 2,
It is placed in -5 DEG C of standing 12h or more in reaction of low temperature plasma device again afterwards, forms uniform colloidal sol, further take out evaporation drying and be changed into
Gel obtains nanogel particle of the size less than 0.1 μm；The Ago-Gel is with agarose and TAE solution with the matter of 2:3
Amount ratio is formulated.
A kind of preparation method of above-mentioned nanogel slant medium, includes the following steps:
A) the preparation of culture medium base fluid: by the slant agar culture medium formula of required purposes, its in addition to agar powder is weighed
His raw material, dissolution are stirred into culture medium base fluid, adjust and arrive required pH；
B the desired amount of agar calculated) the preparation of culture medium: is added into the culture medium base fluid for having regulated pH
Heating stirring adds nanogel particle to being completely dissolved after powder, Ago-Gel, continues heating stirring to being completely dissolved, presses
Preparation method packing, the inclination solidification of slant agar culture medium obtain purpose culture medium；
C) post-process: heat preservation for standby use after sterilization treatment obtains required nanogel slant medium.
The sterilization treatment concrete mode is that chemical sterilization processing mode living carries out, and the mode of autoclave sterilization can be to agarose
Gel generates destruction.
Embodiment 2:
A kind of nanogel slant medium specifically includes following ingredient: grape for the culture of staphylococcus aureus
Sugared 20g, peptone 15g, sodium chloride 5g, beef extract 0.5g, agar 11g, nanogel particle 4g, Ago-Gel 5g；Add steaming
Distilled water is formulated into 1L；
The nanogel particle is the preparation of zinc oxide-titanic oxide nano compound material: zinc chloride is dissolved in ten times
Titania powder continuation is added after reacting 20min in the acid solution of the dehydrated alcohol of volume, in low-temperature plasma reactor
30min is reacted, reaction temperature is -6 DEG C, and it is uniform to solution dispersion to take out sufficiently ultrasonic agitation after completion of the reaction；Adjusting pH is 2-
3, after be placed in 0 DEG C of standing 12h or more in reaction of low temperature plasma device again, form uniform colloidal sol, further take out evaporation drying transformation
For gel, nanogel particle of the size less than 0.1 μm is obtained；The Ago-Gel is with agarose and TAE solution with 2:3
Mass ratio is formulated.
A kind of preparation method of nanogel slant medium is same as Example 1.
Embodiment 3:
A kind of nanogel slant medium specifically includes following ingredient: grape for the culture of staphylococcus aureus
Sugared 20g, peptone 15g, sodium chloride 5g, beef extract 0.5g, agar powder 9g, nanogel particle 5g, Ago-Gel 6g；Add steaming
Distilled water is formulated into 1L；
The nanogel particle is the preparation of zinc oxide-titanic oxide nano compound material: zinc chloride is dissolved in ten times
Titania powder continuation is added after reacting 15min in the acid solution of the dehydrated alcohol of volume, in low-temperature plasma reactor
25min is reacted, reaction temperature is -9 DEG C, and it is uniform to solution dispersion to take out sufficiently ultrasonic agitation after completion of the reaction；Adjusting pH is
2.5, after be placed in -3 DEG C of standings 12h or more in reaction of low temperature plasma device again, form uniform colloidal sol, further take out evaporation drying turn
Become gel, obtains nanogel particle of the size less than 0.1 μm；The Ago-Gel is with agarose and TAE solution with 2:3
Mass ratio be formulated.
A kind of preparation method of nanogel slant medium is same as Example 1.
Comparative example:
A kind of common slant medium specifically includes following ingredient: glucose for the culture of staphylococcus aureus
20g, peptone 15g, sodium chloride 5g, beef extract 0.5g, agar powder 20g plus distilled water are formulated into 1L.
Preparation method is common slant medium preparation method.
Beneficial effect verification experimental verification:
Examination is compared with the common slant medium of the nanogel slant medium of embodiment 1-3 preparation and comparative example
It tests；Staphylococcus aureus is cultivated, measures setting time and observation ramp thickness uniform level.
Every group of three parallel tests, while distribution of culture medium situation is observed, soft or hard degree.It the results are shown in Table 1:
1 embodiment 1-3 of table and control group setting time and ramp thickness situation comparative analysis
It can be seen from the data in Table 1 that the nanogel slant medium of embodiment 1-3 is compared with control group, for training
It is preferable to support control in base setting rate, and slant medium caliper uniformity degree is high, also improves culture medium nutriment
Uniform distribution of forces avoids more so that the best cultivation conditions of bacterium and the accuracy of follow-up test result be effectively ensured
It is secondary to repeat to test, shorten test period, reduce error, is conducive to the levels of precision for improving test, in addition, preparation side of the invention
Method design is rationally, simple to operate.
It should be pointed out that the above specific embodiment is only illustrative of the invention and is not intended to limit the scope of the invention,
After having read the present invention, those skilled in the art fall within power appended by the application to the modification of various equivalent forms of the invention
Benefit requires limited range.
Claims (4)
1. a kind of nanogel slant medium, it is characterised in that received on the basis of common slant agar culture medium with part
Rice gel particles and the agar powder of Ago-Gel substitution part, are specifically calculated first based on mass fraction with nanogel particle
Substitute the agar powder of 20%-30%, then the agar powder with Ago-Gel substitution 25%-35%；Wherein, common slant agar culture medium
Formula are as follows: glucose 20g, peptone 15g, sodium chloride 5g, beef extract 0.5g, agar powder 20g, plus distilled water be formulated into 1L；
The nanogel particle is not higher than 50% in the mass percentage content in addition to cultivating based solvent distilled water；The nanogel
The size of particle is less than 0.1 μm；
The nanogel particle is zinc oxide-titanic oxide nano compound material, preparation method are as follows: zinc oxide is dissolved in ten
In the acid solution of the dehydrated alcohol of times volume, titania powder is added after reacting 10-20min in low-temperature plasma reactor
The reaction was continued 20-30min, reaction temperature are -12~-6 DEG C, and it is equal to solution dispersion to take out sufficiently ultrasonic agitation after completion of the reaction
It is even；Adjusting pH is 2-3, after be placed in -5-0 DEG C of standing 12h or more in reaction of low temperature plasma device again, form uniform colloidal sol, then
It takes out evaporation drying and is changed into gel, obtain nanogel particle.
2. nanogel slant medium according to claim 1, it is characterised in that: the Ago-Gel is with agar
Sugar is formulated with TAE solution with the mass ratio of 2:3.
3. the preparation method of nanogel slant medium as described in claim 1, it is characterised in that include the following steps:
A by the slant agar culture medium formula of required purposes, other originals in addition to agar powder) the preparation of culture medium base fluid: are weighed
Material, dissolution are stirred into culture medium base fluid, adjust and arrive required pH；
B the desired amount of agar powder, the fine jade calculated) the preparation of culture medium: is added into the culture medium base fluid for having regulated pH
Heating stirring adds nanogel particle to being completely dissolved after sepharose, continues heating stirring to being completely dissolved, by inclined-plane
Preparation method packing, the inclination solidification of agar medium obtain purpose culture medium；
C) post-process: heat preservation for standby use after sterilization treatment obtains required nanogel slant medium.
4. the preparation method of nanogel slant medium according to claim 3, it is characterised in that: the sterilization treatment
Concrete mode is that chemical sterilization processing mode living carries out.
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| CN109827814A (en) * | 2019-03-11 | 2019-05-31 | 南京大学 | A kind of sampling film preparation of novel air particulate matter agar and exempt from solvent extraction cell process for exposing |
| CN112514797B (en) * | 2020-12-22 | 2021-06-11 | 甘肃省农业科学院林果花卉研究所 | Peach stock culture medium and use method thereof |
| WO2023080204A1 (en) * | 2021-11-04 | 2023-05-11 | Ｓｙｍｂｉｏｂｅ株式会社 | Culture method for promoting proliferation and carbon fixation of purple photosynthetic bacteria |
| CN114058488B (en) * | 2021-12-15 | 2023-10-13 | 涌源合生科技(深圳)有限公司 | Strain culture device based on enzyme engineering is convenient for coating |
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| CN103898040A (en) * | 2014-03-13 | 2014-07-02 | 浙江大学 | Nano-zinc oxide-based culture medium capable of promoting microbial growth |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103898040A (en) * | 2014-03-13 | 2014-07-02 | 浙江大学 | Nano-zinc oxide-based culture medium capable of promoting microbial growth |
Non-Patent Citations (2)
| Title |
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| 壳聚糖衍生物对Zn(Ⅱ)的吸附行为;丁萍等;《化工学报》;20061130;2652-2656 * |
| 纳米氧化锌的研究进展;辛显双等;《化学研究与应用》;20031031;601-606 * |
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