36 kinds of microbial experiments commonly used in culture
1. Beef paste peptone medium (for bacterial culture)
Beef paste 3g, peptone 10g, NaCl 5g, water 1000mL, pH 7.4~ 7.6.
2. Gao's No. 1 medium (for actinomycete culture)
Soluble starch 20g, KNO3 1g, NaCl 0.5g, K2HPO4? 3H2O 0.5g, MgSO4? 7H2O0.5g, FeSO4? 7H2O0.01g, water 1000mL, pH 7.4~ 7.6. Note when preparing: Soluble starch should be mixed with cold water first and then added to the abovemediumIn.
3. Martin's medium (used to isolate fungi from soil)
K2HPO41g, MgSO4? 7H2O0.5g, peptone 5g, glucose 10g, 1/3000 Bengal red aqueous solution 100mL, water 900mL, natural pH, 121 ℃ damp heat sterilization for 30min. Add streptomycin (streptomycin content is 30μg/mL) when the medium is melted and cooled at 55-60 ° C.
4. Potato medium (PDA medium) (for mold or yeast culture)
Potato (peeled) 200g, sucrose (or glucose) 20g, water 1000mL, the preparation method is as follows:
Peel the horse bell department, cut it into small pieces of about 2cm2, put it in a 1500mL beaker and boil for 30min, pay attention to stirring with a glass rod to prevent the bottom from being pasted, then filter it with double-layer gauze, take the filtrate and add sugar, and then make up to 1000mL, natural pH, sucrose for mold and glucose for yeast.
5. Chasteur medium (sucrose sodium nitrate medium) (for mold culture)
蔗糖30g，NaNO3 2g，K2HPO4 1g，MgSO4?7H2O0.5g，KCl 0.5g，FeSO4?7H2O0.1g，水1000mL，pH7.0～7.2。
6. Hayflik medium (for Mycoplasma culture)
Beef heart digestive solution (or extract) 1000mL, peptone 10g, NaCl 5g, agar 15g, pH 7.8~ 8.0, sub-packaged 70mL per bottle, 121 ℃ damp heat sterilization 15min, to be cooled to about 80 ℃, every 70mL was added horse serum 20mL, 25% fresh yeast extract 10mL, 15 thallium acetate aqueous solution 2.5mL, penicillin G potassium salt aqueous solution (200,000 units or more) 0.5mL, after mixing the above poured plate.
Attention: Thallium acetate is an extremely toxic drug and requires special attention to safe operation.
7. McCLary medium (sodium acetate medium)
Glucose 0.1g, KCl 0.18g, yeast cream 0.25g, sodium acetate 0.82g, agar 1.5g, distilled water l00mL. After dissolving, separate test tubes, 1l5 ℃ damp heat sterilization for 15min.
8. Glucose peptone water medium (for V.P. reaction and methyl red test)
Peptone 0.5g, glucose 0.5g, K2HPO4 0.2g, water 100mL, pH 7.2, 1l5 ℃ humid heat sterilization for 20min.
9. Peptone water medium (for indole test)
Peptone 10g, NaCl 5g, water 1000mL, pH 7.2~ 7.4, 121 ℃ moist heat sterilization 20min.
10. Sugar fermentation medium (for bacterial sugar fermentation test)
Peptone 0.2g, NaCl 0.5g, K2HPO4 0.02g, water 100mL, bromothymol blue (1% aqueous solution) 0.3mL, sugar lg. Weigh the peptone and NaCl respectively and dissolve them in hot water, adjust the pH to 7.4, then add bromothymol blue (first dissolve with a small amount of 95% ethanol, then add water to prepare a 1% aqueous solution), add sugar, pack test tubes separately, the loading is 4~ 5cm high, and pour them into a Duchenne tube (the nozzle is down, and the tube is filled with culture solution). Sterilize at 115 ° C with humid heat for 20min. When sterilizing, pay attention to appropriately extend the boiling time, and try to drain the cold air so that no bubbles remain in the Duchenne tube. Commonly used sugars, such as glucose, sucrose, mannose, maltose, lactose, galactose, etc. (the amount of the latter two sugars is often increased to 1.5%).
11. RCM medium (enhanced Clostridium medium), (for anaerobic culture)
Yeast cream 3g, l0g beef paste, peptone 10g, soluble starch lg, glucose 5g, 0.5g cysteine hydrochloride, NaCl 3g, NaAc 3g, water l000mL, pH 8.5, blade azure 3mg/L, l2l ℃ damp heat sterilization 30min.
12. TYA medium (for anaerobic culture)
Glucose 40g, beef paste 2g, yeast paste 2g, tryptone (bacto-typetone) 6g, ammonium acetate 3g, KH2PO4 0.5g, MgSO4? 7H2O0.2g, FeSO4? 7H2O0.01g, water 1000mL, pH 6.5, 121 ℃ humid heat sterilization for 30min.
13. Corn mash medium (for anaerobic culture)
Corn flour 65g, tap water 1000mL, mix well, cook 10min into a paste, natural pH, 121 ℃ humid heat sterilization 30min.
14. Neutral red medium (for anaerobic culture)
Glucose 40g, tryptone 6g, yeast paste 2g, beef paste 2g, ammonium acetate 3g, KH2PO4 5g, neutral red 0.2g, MgSO4? 7H2O0.2g, FeSO4? 7H2O0.01g, water 1000mL, pH 6.2, 121 ℃ humid heat sterilization for 30min.
15. CaCO3 gelatin wort medium (for anaerobic culture)
Wort (6 Baume) 1000mL, water 1000mL, CaCO310g, gelatin 10g, pH 6.8, 121 ℃ humid heat sterilization 30min.
16.BCG milk medium (for lactic acid fermentation)
(A) Solution:
100G defatted milk powder, 500mL water, add 1.6% bromocresol green (B.C.G) ethanol solution 1mL, sterilize at 80 ℃ for 20min.
(B) Solution:
Yeast cream 10g, water 500mL, agar 20g, pH 6.8, 121 ℃ moist heat sterilization 20min. Mix (A) and (B) solutions evenly while hot in aseptic operation and then pour the plate.
17. Lactic acid bacteria medium (for lactic acid fermentation)
Beef paste 5g, yeast paste 5g, peptone 10g, glucose 10g, lactose 5g, NaCl 5g, water 1000mL, pH 6.8, 121 ℃ humid heat sterilization 20min.
18. Alcohol fermentation medium (for alcoholic fermentation)
蔗糖10g,MgSO4?7H2O 0.5g,NH4NO30.5g,20%豆芽汁2mL，KH2PO4 0.5g，水100mL，自然pH。
19. Kosov medium (for Leptospira culture)
Quality peptone 0.4g, NaCl 0.7g, KCl 0.02g, NaHCO3 0.01g, CaCl 0.02g, KH2PO40.09g, NaH2PO40.48g, distilled water 500mL, sterile rabbit serum 40mL.
Production method:
The remaining ingredients except rabbit serum were mixed, heated and dissolved, pH was adjusted to 7.2, and humid heat was sterilized at 121 ° C for 20 minutes. After cooling, sterile rabbit serum was added to prepare an 8% serum solution, and then sub-packed into test tubes (5~ 10 mL/tube). 56 ° C water bath was inactivated for 1 h before standby.
20. Sprouts juice medium
500G soybean sprouts, add 1000mL of water, boil for 1h, filter to make up for water, and store for later use after humid heat sterilization at 121 ° C, which is 50% bean sprouts juice;
For bacterial culture:
10% bean sprout juice 200mL, glucose (or sucrose) 50g, water 800mL, pH 7.2~ 7.4.
For mold or yeast cultures:
10% bean sprout juice 200mL, sugar 50g, water 800mL, natural pH. Sucrose for mold, glucose for yeast.
21.LB (Luria-Bertani) medium (bacterial culture, often used in molecular biology)
Double distilled water 950mL, tryptone l0g, NaCl l0g, yeast extract (bacto- yeast extract) 5g, with lmol/L NaOH (about l mL) to adjust the pH to 7.0, add double distilled water to a total volume of 1L, 121 ℃ moist heat sterilization 30min.
Ampicillin-containing LB medium:
After sterilizing the LB medium, add antibiotics at about 50 ° C until the final concentration is 80-100 mg/L.
22. Double red sodium sulfite medium (Endo's medium), (for the determination of coliform in water) peptone 10g, beef extract 5g, yeast extract 5g, agar 20g, lactose 10g, K2HPO40.5g, anhydrous sodium sulfite 5g, 5% basic double red ethanol solution 20mL, distilled water 1000mL.
Production process:
First add peptone, beef extract, yeast extract and agar to 900mL of water, heat and dissolve, then add K2PO4, add water to l000mL after dissolution, and adjust pH to 7.2~ 7.4. Then add lactose, mix well and dissolve, sterilize under moist heat at 115 ° C for 20 minutes. Then weigh sodium sulfite into a sterile empty test tube, dissolve it with a little sterile water, boil it in a water bath for 10 minutes, and immediately add it dropwise to 20mL of 5% alkaline reredding ethanol solution until dark red turns to light pink. Add this mixture to the above sterilized medium and still keep it in a melted state. Immediately pour the plate after mixing, and store it in the refrigerator for later use after solidification. If the color changes from light red to dark red, it cannot be used again.
23. Lactose peptone semi-solid medium (for the determination of coliform in water)
Peptone 10g, beef extract 5g, yeast paste 5g, lactose 10g, agar 5g, distilled water 1000mL, pH 7.2~ 7.4, sub-packed test tubes (l0mL/tube), 115 ℃ humid heat sterilization 20min.
24. Lactose peptone medium (used for multi-tube fermentation to detect coliform bacteria in water)
Peptone 10g, beef paste 3g, lactose 5g, NaCl 5g, distilled water l000mL, 1.6% bromocresol violet ethanol solution lmL. Adjust pH to 7.2, sub-pack test tubes (l0mL/tube), and put into inverted Duchenne tubules, 15 ℃ humid heat sterilization for 20min.
25. Triple concentrated lactose peptone medium (for the determination of coliform in water)
The nutrients in the lactose peptone medium were added to 1000 mL of water to expand by 3 times, and the preparation method was the same as above. They were sub-packaged in test tubes with inverted Duchenne tubules, each tube was 5 mL, and the humid heat at 115 ° C was sterilized for 20 minutes.
26. Eosin methylene blue medium (EMB medium) (for coliform determination and bacterial transduction in water)
Peptone l0g, lactose 10g, K2HPO4 2g, agar 25g, 2%/eosin Y (eosin) aqueous solution 20mL, 0.5% methylene blue (methylene blue) aqueous solution l3mL, pH 7.4.
Production process:
First, mix peptone, lactose, K2HPO4 and agar, heat and dissolve, adjust the pH to 7.4, 1l5 ℃ moist heat sterilization for 20min, then add the sterilized eosin liquid and methylene blue liquid, mix well to prevent bubbles. Wait for the medium to cool to about 50 ℃ and pour the plate. If the medium is too hot, it will produce too much coagulated water, which can be stored upside down in the refrigerator after the plate solidifies. Galactose is used instead of lactose in the bacterial transduction experiment, and the rest of the ingredients remain unchanged.
27. Double broth medium (for bacterial transduction)
Beef paste 6g, peptone 20g, NaCl 10g, water 1000mL, pH 7.4~ 7.6.
28. Semi-solid agar (for bacterial transduction) agar 1g, water 100mL, 121 ℃ moist heat sterilization for 30 minutes.
29. Soybean cake bevel medium (for screening protease-producing mold strains)
Add water to 100g of soybean cake 5 to 6 times, cook 100mL of filter juice, add KH2PO4 0.1%, MgSO4 0.05%, (NH4) 2SO4 0.05%, soluble starch 2%, pH 6, agar 2% to 2.5%.
30. Casein medium (for protease strain screening)
Prepare solution A and solution B, respectively.
A liquid:
Weigh 1.07g of Na2HPO4? 7H2O. 4g of casein, add an appropriate amount of distilled water, and heat to dissolve.
Liquid B:
Weigh KH2PO4 0.36g, add water to dissolve. A, B solution mixed, add 0.3 mL casein hydrolysate, add agar 20g, and finally with distilled water bandwidth evaluation to 1000mL.
Preparation of casein hydrolysate:
1G casein is dissolved in alkaline buffer, add 1% Bacillus subtilis protease 25mL with water to 100mL, and hydrolyze at 30 ° C for 1 h. When used to prepare medium, the dosage is 1000mL, and more than 100mL of hydrolytic solution is added to the medium.
31. Bacterial basic medium (for screening for nutrient deficiencies)
Na2HPO4?7H2O 1g，MgSO4?7H2O0.2g，葡萄糖5g，NaCl 5g，K2HPO4lg，水l000mL，pH7.0，1l5℃湿热灭菌30min。
32. YEPD medium (for yeast protoplast fusion)
Yeast powder 10g, peptone 20g,glucose20G, distilled water 1000mL, pH 6.0, 115 ℃ moist heat sterilization 20min.
33. YEPD hyperosmotic medium (for yeast protoplast fusion)
Add 0.6mol/L of NaCL, 3% agar to YEPD medium.
34. YNB basic medium (for yeast protoplast fusion)
Yeast nitrogen base 0.67% (YNB, without amino acids, Difco), 2% glucose, 3% agar, pH 6.2.
Another recipe is:
Glucose 10g (NH4) 2SO4 1g, K2HPO40.125g, KHPO4 0.875g, KI 0.0001g, MgSO4? 7H2O0.5g, CaCl2? 2H2O0.lg, NaCl0.1g, dimensional element mother liquor lmL, vitamin mother liquor lmL (mother liquor are conventionally prepared), water 1000mL, pH 5.8~ 6.0.
35. YNB hyperosmotic basic medium (for protoplast fusion)
Add 0.6mol/L NaCl to YNB basal medium.
36. Phenol red semi-solid columnar medium (used to check the relationship between oxygen and bacterial growth)
Peptone 1g, glucose 10g, corn syrup 10g, agar 7g, water 1000mL, pH 7.2. After adjusting the pH, add a few drops of 1.6% phenol red solution until the medium turns dark red, sub-packed in large test tubes, the loading is about 1/2 of the height of the test tube, and sterilized at 1l5 ° C for 20min. Bacteria use glucose growth in this medium to produce acid, so that phenol red changes from red to yellow. Bacteria growing in different parts can change the color of the corresponding parts of the medium, but pay attention to the culture time is too long, and the acid can spread so that the result cannot be correctly judged.
The above various media can be formulated into a solid or semi-solid state, and only the amount of agar can be changed. The former is 1.5% to 2.0%, and the latter is 0.3% to 0.8%.
