CN106011029A - Microbial plate culture method - Google Patents
Microbial plate culture method Download PDFInfo
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- CN106011029A CN106011029A CN201610597324.7A CN201610597324A CN106011029A CN 106011029 A CN106011029 A CN 106011029A CN 201610597324 A CN201610597324 A CN 201610597324A CN 106011029 A CN106011029 A CN 106011029A
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- culture
- bacterium colony
- active sludge
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- Wood Science & Technology (AREA)
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
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Abstract
The invention discloses a microbial plate culture method. The microbial plate culture method comprises the steps of plate culture medium preparation, active sludge mixed liquor preparation, culture medium coating, bacterial colony culture and bacterial colony separation. The microbial plate culture method is easy to operate, has no requirement on colonial morphology of bacteria, and is convenient for observing colony characteristics and colony counting; by adopting the method, the requirement on the plate culture medium is lowered, and production and scientific research work are ensured.
Description
Technical field
The present invention relates to field of microbial culture technology, particularly to a kind of microorganism flat board cultural method.
Background technology
In order to observe and study bacterium colony situation, the most generally employing following two microbial culture method:
1, plate streaking partition method
Different cells in microorganism mixed in together or same micropopulation are inoculated ring at flat board
Media surface, is diluted by sectional streak and obtains the more individual cells being independently distributed, raw after cultivating
Length is multiplied into single bacterium colony.
2, dilution spread flat band method
Bacterium solution is carried out a series of gradient dilution, then different dilution bacterium solution is respectively coated agar
The surface of solid medium is cultivated, in the sufficiently high bacterium solution of dilution factor, and the microorganism flocked together
Individual cells will be dispersed into, it is thus possible to form single bacterium colony in media surface.
Although above two method can realize observing the purpose of colony characteristics, but plate streaking partition method is not
Can be to colony counting, and dilution spread flat band method incubation is complicated, and the requirement to flat board is higher.
Summary of the invention
For solving above-mentioned technical problem, the invention discloses a kind of microorganism flat board cultural method, including following
Step:
A, prepare plating medium
The heating of sodium chloride sucrose agar culture medium is dissolved and is cooled to 65 DEG C, cultivates to sodium chloride sucrose agar
Base adds clindamycin hydrochloride solution 3～5ml mix homogeneously, then takes the culture medium solution of 20ml mix homogeneously
Pour culture dish into, be shaken gently for culture dish, make culture medium solution be evenly distributed on bottom culture dish, wait to cultivate
Plating medium is i.e. obtained after based sols solidification.
B, prepare active sludge intermixture
Weigh activated sludge soil sample 15g, put in Sheng 100ml sterilized water the conical flask with bead,
Shaking 15～20min mix homogeneously, therefrom draw the addition of 1ml soil supension with a 1ml aseptic straw and fill
The Boiling tube of 9ml sterilized water fully mixes, makes active sludge intermixture.
C, culture medium are coated with
From active sludge intermixture, draw 0.2ml, drop in the middle position on plating medium surface, with aseptic
Glass is coated with rod by active sludge intermixture along concentric circular direction outward expansion lightly, is allowed to be evenly distributed, so
After at room temperature stand 5～l0min, make active sludge intermixture immerse culture medium.
D, bacterium colony are cultivated
Plating medium is inverted in the greenhouse of 28 DEG C cultivation 3～5 days, forms bacterium colony.
E, bacterium colony separate
The single bacterium colony grown after cultivating is observed, afterwards according to features such as its size, color, shapes
It is inoculated on plating medium according to the bacterium colony a little cell of different characteristic picking, is placed in the hot-house culture of 28 DEG C；
If being found to have miscellaneous bacteria, need to carry out again separating, purification, until obtaining pure culture.
The invention has the beneficial effects as follows:
The microorganism flat board cultural method of the present invention is simple to operate, the form not requirement to bacterial clump, fall
The low requirement to plating medium, and be easy to observe colony characteristics and colony counting, for producing and section
Grind work and provide basic guarantee.
Accompanying drawing explanation
For the technical scheme being illustrated more clearly that in the embodiment of the present invention, below will be to required in embodiment
The accompanying drawing used is briefly described, it should be apparent that, the accompanying drawing in describing below is only the one of the present invention
A little embodiments, for those of ordinary skill in the art, on the premise of not paying creative work,
Other accompanying drawing can also be obtained according to these accompanying drawings.
Fig. 1 is the process sequence diagram of microorganism flat board cultural method of the present invention.
Detailed description of the invention
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clearly
Chu, be fully described by, it is clear that described embodiment be only a part of embodiment of the present invention rather than
Whole embodiments.
As it is shown in figure 1, the microorganism flat board cultural method of the present invention, comprise the following steps:
A, prepare plating medium
The heating of sodium chloride sucrose agar culture medium is dissolved and is cooled to 65 DEG C, cultivates to sodium chloride sucrose agar
Base adds clindamycin hydrochloride solution 3～5ml mix homogeneously, then takes the culture medium solution of 20ml mix homogeneously
Pour culture dish into, be shaken gently for culture dish, make culture medium solution be evenly distributed on bottom culture dish, wait to cultivate
Plating medium is i.e. obtained after based sols solidification.
B, prepare active sludge intermixture
Weigh activated sludge soil sample 15g, put in Sheng 100ml sterilized water the conical flask with bead,
Shaking 15～20min mix homogeneously, therefrom draw the addition of 1ml soil supension with a 1ml aseptic straw and fill
The Boiling tube of 9ml sterilized water fully mixes, makes active sludge intermixture.
C, culture medium are coated with
From active sludge intermixture, draw 0.2ml, drop in the middle position on plating medium surface, with aseptic
Glass is coated with rod by active sludge intermixture along concentric circular direction outward expansion lightly, is allowed to be evenly distributed, so
After at room temperature stand 5～l0min, make active sludge intermixture immerse culture medium.
D, bacterium colony are cultivated
Plating medium is inverted in the greenhouse of 28 DEG C cultivation 3～5 days, forms bacterium colony.
E, bacterium colony separate
The single bacterium colony grown after cultivating is observed, afterwards according to features such as its size, color, shapes
It is inoculated on plating medium according to the bacterium colony a little cell of different characteristic picking, is placed in the hot-house culture of 28 DEG C；
If being found to have miscellaneous bacteria, need to carry out again separating, purification, until obtaining pure culture.
The microorganism flat board cultural method of the present invention is simple to operate, the form not requirement to bacterial clump, fall
The low requirement to plating medium, and be easy to observe colony characteristics and colony counting, for producing and section
Grind work and provide basic guarantee.
The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, all at this
Within bright spirit and principle, any modification, equivalent substitution and improvement etc. made, should be included in this
Within bright protection domain.
Claims (1)
1. microorganism flat board cultural method, it is characterised in that comprise the following steps:
A, prepare plating medium
The heating of sodium chloride sucrose agar culture medium is dissolved and is cooled to 65 DEG C, cultivates to sodium chloride sucrose agar
Base adds clindamycin hydrochloride solution 3～5ml mix homogeneously, then takes the culture medium solution of 20ml mix homogeneously
Pour culture dish into, be shaken gently for culture dish, make culture medium solution be evenly distributed on bottom culture dish, wait to cultivate
Plating medium is i.e. obtained after based sols solidification.
B, prepare active sludge intermixture
Weigh activated sludge soil sample 15g, put in Sheng 100ml sterilized water the conical flask with bead,
Shaking 15～20min mix homogeneously, therefrom draw the addition of 1ml soil supension with a 1ml aseptic straw and fill
The Boiling tube of 9ml sterilized water fully mixes, makes active sludge intermixture.
C, culture medium are coated with
From active sludge intermixture, draw 0.2ml, drop in the middle position on plating medium surface, with aseptic
Glass is coated with rod by active sludge intermixture along concentric circular direction outward expansion lightly, is allowed to be evenly distributed, so
After at room temperature stand 5～l0min, make active sludge intermixture immerse culture medium.
D, bacterium colony are cultivated
Plating medium is inverted in the greenhouse of 28 DEG C cultivation 3～5 days, forms bacterium colony.
E, bacterium colony separate
The single bacterium colony grown after cultivating is observed, afterwards according to features such as its size, color, shapes
It is inoculated on plating medium according to the bacterium colony a little cell of different characteristic picking, is placed in the hot-house culture of 28 DEG C；
If being found to have miscellaneous bacteria, need to carry out again separating, purification, until obtaining pure culture.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610597324.7A CN106011029A (en) | 2016-07-27 | 2016-07-27 | Microbial plate culture method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610597324.7A CN106011029A (en) | 2016-07-27 | 2016-07-27 | Microbial plate culture method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106011029A true CN106011029A (en) | 2016-10-12 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201610597324.7A Pending CN106011029A (en) | 2016-07-27 | 2016-07-27 | Microbial plate culture method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111735919A (en) * | 2020-07-02 | 2020-10-02 | 浙江莫干山食业有限公司 | Preserved fruit microorganism detection technology |
| CN114009450A (en) * | 2021-11-15 | 2022-02-08 | 郭云 | Preparation method of microbial preparation prepared by using Streptomyces rochei |
| WO2023279490A1 (en) * | 2021-07-07 | 2023-01-12 | 深圳先进技术研究院 | Colony coating process and colony culturing process |
-
2016
- 2016-07-27 CN CN201610597324.7A patent/CN106011029A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 张勇主编: "《生物样本库建设与实践》", 31 October 2013 * |
| 赵开弘主编: "《环境微生物学》", 31 July 2009 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111735919A (en) * | 2020-07-02 | 2020-10-02 | 浙江莫干山食业有限公司 | Preserved fruit microorganism detection technology |
| WO2023279490A1 (en) * | 2021-07-07 | 2023-01-12 | 深圳先进技术研究院 | Colony coating process and colony culturing process |
| CN114009450A (en) * | 2021-11-15 | 2022-02-08 | 郭云 | Preparation method of microbial preparation prepared by using Streptomyces rochei |
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Application publication date: 20161012 |
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