LB solid culture medium preparing method

A technology of solid medium and petri dish, which is applied in the direction of bacteria, can solve the problems of weak ability to absorb bacterial liquid, consume a long time, delay the test process, etc., and achieve the goal of promoting full absorption, improving test efficiency and prolonging storage time Effect

Inactive Publication Date: 2016-10-12
SINOBIOWAY CELL THERAPY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The LB solid medium prepared by this method, because the medium cannot be completely solidified, after the plate is stored upside down, more condensed water will appear on the lid of the petri dish, resulting in inconvenience in use, prone to bacterial contamination, and short storage life; Moreover, the ability of the solid medium to absorb the bacterial solution is weak. When doing experiments, it will take a long time to absorb the bacterial solution, which will delay the test process.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A preparation method for LB solid medium, comprising the steps:

[0024] S1. Add tryptone, yeast extract, and sodium chloride into water to dissolve, adjust the pH to 7.2 with aqueous sodium hydroxide solution, add agar powder and mix evenly, and obtain solution A at constant volume;

[0025] S2. Take the solution A obtained in S1, autoclave it, cool to 50°C, add antibiotics, and shake well to obtain solution B;

[0026] S3. Add the solution B obtained in S2 into a petri dish, adjust the wind speed to 1000r / min, blow air for 70min, and seal the petri dish with a sterile parafilm to obtain LB solid medium.

Embodiment 2

[0028] A preparation method for LB solid medium, comprising the steps:

[0029] S1. Add tryptone, yeast extract, and sodium chloride into water to dissolve, adjust the pH to 7.3 with a 0.5 mol / l sodium hydroxide aqueous solution, add agar powder and mix well, and obtain solution A at constant volume, wherein, solution A The concentration of tryptone in medium is 9g / l, the concentration of yeast extract is 6g / l, the concentration of sodium chloride is 9g / l, and the concentration of agar powder is 16g / l;

[0030] S2. Take the solution A obtained in S1, autoclave it for 20 minutes, cool it to 55°C, transfer it to an ultra-clean workbench, add antibiotics, and shake well to obtain solution B;

[0031] S3. On the ultra-clean bench, add the solution B obtained in S2 into a petri dish, adjust the wind speed to 800r / min, blow for 80 minutes, seal the petri dish with a sterile sealing film, and store it at 2°C to obtain LB solid medium.

Embodiment 3

[0033] A preparation method for LB solid medium, comprising the steps:

[0034] S1. Add tryptone, yeast extract, and sodium chloride into water to dissolve, adjust the pH to 7.1 with a 1.5mol / l sodium hydroxide aqueous solution, add agar powder and mix well, and obtain solution A at constant volume, wherein, solution A The concentration of tryptone in medium is 11g / l, the concentration of yeast extract is 4g / l, the concentration of sodium chloride is 11g / l, and the concentration of agar powder is 14g / l;

[0035] S2. Take the solution A obtained in S1, autoclave for 30 minutes, cool to 45°C, transfer to a clean bench, add antibiotics, and shake well to obtain solution B;

[0036] S3. Add the solution B obtained in S2 into a petri dish on the ultra-clean workbench, adjust the wind speed to 1200r / min, blow for 60 minutes, seal the petri dish with a sterile sealing film, and store it at 4°C to obtain LB solid medium.

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PUM

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Abstract

The invention discloses an LB solid culture medium preparing method. The method comprises the steps of S1, adding tryptone, yeast extract and sodium chloride to water to be dissolved, adjusting pH to be 7.1-7.3 with sodium hydroxide aqueous solution, adding agar powder, conducting uniform mixing, and conducting volume metering to obtain solution A; S2, conducting high-pressure sterilization on the solution A obtained from S1, reducing temperature to 45-55 DEG C, adding antibiotics, and conducting uniform shaking to obtain solution B; S3, adding the solution B obtained from S2 into a culture dish, regulating air speed to be 800-1200 r/min, conducting blowing for 60-80 min, and sealing the culture dish with a sterile sealing film to obtain the LB solid culture medium. By the adoption of the method, generation of condensate water during LB solid culture medium preparation is avoided, the situation that the culture medium is polluted by bacteria is avoided, waste is reduced, the storage time of the culture medium is prolonged, the bacterial solution absorption capacity of the culture medium is improved greatly, test time is shortened, and test efficiency is improved.

Description

technical field [0001] The invention relates to the technical field of medium preparation, in particular to a preparation method of LB solid medium. Background technique [0002] LB medium is the name of a medium. In biochemical molecular experiments, this medium is generally used to pre-cultivate bacteria to multiply the bacteria and meet the requirements of use. It can be divided into liquid medium and solid medium. [0003] At present, the commonly used methods for preparing LB solid medium are: preparation of medium solution, sterilization, plating, solidification, and upside-down storage. The LB solid medium prepared by this method, because the medium cannot be completely solidified, after the plate is stored upside down, more condensed water will appear on the lid of the petri dish, resulting in inconvenient use, prone to bacterial contamination, and short storage life; Moreover, the ability of the solid medium to absorb the bacterial solution is relatively weak. When...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20
CPCC12N1/20
Inventor 刘振云许国贞
Owner SINOBIOWAY CELL THERAPY CO LTD
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