Experiments on the preparation of Martin's medium
Experiments on the preparation of Martin's medium
Martin's medium is a selective medium used to isolate fungi. This medium is composed of glucose, peptone, K2HPO4, MgS04-7H2O, Bengal red (Rose Bengal), and streptomycin, etc., where glucose is mainly used as a carbon source, peptone is mainly used as a nitrogen source, and K2HPO4 and MgS04-7H2O are used as inorganic salts to provide the microorganisms with potassium, phosphorus, and magnesium ions.
Operation method
Preparation of Martin's medium
Principle
Martin's medium is a selective medium used to isolate fungi. This medium is composed of glucose, peptone, K2HPO4, MgS04-7H2O, Bengal red (Rose Bengal) and streptomycin, etc., in which glucose is mainly used as a carbon source, peptone is mainly used as a nitrogen source, and K2HPO4 and MgS04-7H2O are used as inorganic salts to provide microorganisms with potassium, phosphorus and magnesium ions. This kind of medium is characterized by the fact that the addition of Bengal red and streptomycin in the medium can effectively inhibit the growth of bacteria and actinomycetes, while it has no inhibitory effect on the fungi, thus the fungi can get the advantageous growth on this kind of medium, so as to achieve the purpose of isolation of fungi.
Materials and Instruments
Glucose Peptone K2HPO4 MgS04-7H2O Bengal red Streptomycin Agar Move I. Martin's medium formulation For more product details, please visit Aladdin Scientific website.
Test tubes Triangular flasks Beakers Measuring cylinders Glass rods Medium dispensers Scales Horny horn spoons Autoclaves
This culture solution 1,000 ml plus 1% Bengal red aqueous solution 3.3 ml.
At the time of administration, 0.3 ml of 1% streptomycin was added aseptically to 100 ml of culture medium to give a final concentration of 30 μg/ml.
II. Equipment
1. Solution or reagent
K2HPO4, MgS04-7H2O, peptone, glucose, agar, Bengal red (1% aqueous solution), streptomycin (1% aqueous solution).2. Instruments or other appliances
Test tubes, triangular flasks, beakers, measuring cylinders, glass rods, medium dispensers, scales, crookspoons, autoclaves, etc.
III. Operational steps
1. Weighing and solubilization
According to the formula of the medium, weigh each component accurately and dissolve each component sequentially in less than the required amount of water. After the components are completely dissolved, make up the water to the required volume, and then formulate Bengal red into a 1% solution, add 3.3 ml of 1% Bengal red solution to 1,000 ml of medium, mix well, and then add agar and heat to dissolve it (the method is the same as that of the preparation experiment of Martin's medium ).
2. Dispensing, corking, wrapping, sterilization, sterility checks
with respect to Martin's medium preparation experiment Same.3. Addition of streptomycin
Streptomycin was prepared as a 1% solution, and 0.3 ml of 1% streptomycin solution was added to 100 ml of medium to make 30 μg of streptomycin per ml of medium.Since streptomycin is easily decomposed by heating, when using it, dissolve the medium and wait until the temperature drops to 45-55 ℃ before adding it.