常用微生物培养基配方大全
发布时间：
2022-11-16
作者：
常用微生物培养基配方大全
（1）分离细菌时，在培养基中加入浓度为50U/m；（2）分离放线菌时，在样品中加入0.05%十二烷；（3）分离霉菌和酵母菌时，在培养基中加入青霉素、；（4）分离根霉和毛霉时，由于这些微生物的菌丝易蔓；一般用于分离单一目的微生物的培养基中均含有抑制其；1、细菌培养基；配方一牛肉膏琼脂培养基；牛肉膏0.3克，蛋白胨1.0克，氯化钠0.5克，；水1000毫升；在烧
（1）分离细菌时，在培养基中加入浓度为50U/ml制霉菌素，可以抑制霉菌和酵母菌的生长。
（2）分离放线菌时，在样品中加入0.05%十二烷基磺酸钠（SDS）不仅可以抑制细菌的生长，还能激活放线菌孢子的萌发。加入氟哌酸（5mg/L）+制霉菌素（50mg/L）+青霉素（0.8mg/L）也可以有效地抑制细菌和真菌，而不影响放线菌的生长。
（3）分离霉菌和酵母菌时，在培养基中加入青霉素、链霉素和四环素各30U/ml，可以抑制细菌和放线菌生长。
（4）分离根霉和毛霉时，由于这些微生物的菌丝易蔓延成片，难以得到纯化的菌落，通常在培养基中添加0.1%去氧胆酸钠或山梨醇防止菌丝蔓延，使菌落长得小而紧密。
一般用于分离单一目的微生物的培养基中均含有抑制其他微生物的抑制剂，这些专用的抑制剂在小型实验室配制较麻烦，现已有成品供应，只要直接加入基础培养基即可。如氨苄西林，主要用于分离亲水气单孢菌。
1、细菌培养基
配方一 牛肉膏琼脂培养基
牛肉膏0.3克 ，蛋白胨1.0克，氯化钠 0.5克，琼脂 1.5克，
水 1000毫升
在烧杯内加水100毫升，放入牛肉膏、蛋白胨和氯化钠，用蜡笔在烧杯外作上记号后，放在火上加热。待烧杯内各组分溶解后，加入琼脂，不断搅拌以免粘底。等琼脂完全溶解后补足失水，用10%盐酸或10%的氢氧化钠调整pH值到7.2～7.6,分装在各个试管里，加棉花塞，用高压蒸汽灭菌30分钟。
配方二 马铃薯培养基
取新鲜牛心（除去脂肪和血管）250克，用刀细细剁成肉末后，加入500毫升蒸馏水和5克蛋白胨。在烧杯上做好记号，煮沸，转用文火炖2小时。过滤，滤出的肉末干燥处理，滤液pH值调到7.5左右。每支试管内加入10毫升肉汤和少量碎末状的干牛心，灭菌，备用。
配方四 根瘤菌培养基
葡萄糖 10克 磷酸氢二钾 0.5克
碳酸钙 3克 硫酸镁 0.2克
酵母粉 0.4克 琼脂 20克
水 1000毫升 1%结晶紫溶液 1毫升
先把琼脂加水煮沸溶解，然后分别加入其他组分，搅拌使溶解后，分装，灭菌，备用。
2、放线菌培养基
配方一 淀粉琼脂培养基（高氏培养基）
可溶性淀粉 2克 硝酸钾 0.1克
磷酸氢二钾 0.05克 氯化钠 0.05克
硫酸镁 0.05克 硫酸亚铁 0.001克
琼脂 2克 水 1000毫升
先把淀粉放在烧杯里，用5毫升水调成糊状后，倒入95毫升水，搅匀后加入其他药品，使它溶解。在烧杯外做好记号，加热到煮沸时加入琼脂，不停搅拌，待琼脂完全溶解后，补足失水。调整pH值到7.2～7.4，分装后灭菌，备用。
配方二 面粉琼脂培养基
面粉 60克 琼脂 20克
水 1000毫升
把面粉用水调成糊状，加水到500毫升，放在文火上煮30分钟。另取500毫升水，放入琼脂，加热煮沸到溶解后，把两液调匀，补充水分，调整pH值到7.4，分装，灭菌，备用。
3、真菌培养基
配方一 萨市（Sabouraud’s）培养基
蛋白胨 10克 琼脂 20克
麦芽糖 40克 水 1000毫升
先把蛋白胨、琼脂加水后，加热，不断搅拌，待琼脂溶解后，加入40克麦芽糖（或葡萄糖），搅拌，使它溶解，然后分装，灭菌，备用。
本培养菌是培养许多种类真菌所常用的。
配方二 马铃薯糖琼脂培养基
把马铃薯洗净去皮，取200克切成小块，加水1000毫升，煮沸半小时后，补足水分。在滤液中加入10克琼脂，煮沸溶解后加糖20克（用于培养霉菌的加入蔗糖，用于培养酵母菌的加入葡萄糖），补足水分，分装，灭菌，备用。
把这培养基的pH值调到7.2～7.4，配方中的糖，如用葡萄糖还可用来培养放线菌和芽孢杆菌。
配方三 黄豆芽汁培养基
黄豆芽 100克 琼脂 15克
葡萄糖 20克 水 1000毫升
洗净黄豆芽，加水煮沸30分钟。用纱布过滤，滤液中加入琼脂，加热溶解后放入糖，搅拌使它溶解，补足水分到1000毫升，分装，灭菌，备用。
把这培养基的pH值调到7.2～7.4，可用来培养细菌和放线菌。
配方四 豌豆琼脂培养基
豌豆 80粒 琼脂 5克
水 200毫升
取80粒干豌豆加水，煮沸1小时，用纱布过滤后，在滤液中加入琼脂，煮沸到溶解，分装，灭菌，备用。
4、食用菌菌种培养基
配方一 马铃薯—蔗糖--琼脂培养基
20%马铃薯煮汁 1000毫升
蔗糖 20克 琼脂 18克
把马铃薯洗净去皮后，切成小块。称取马铃薯小块200克，加水1000毫升，煮沸20分钟后，过滤。在滤汁中补足水分到1000毫升，即成20%马铃薯煮汁。在马铃薯煮汁中加入琼脂和蔗糖，煮沸，使它溶解后，补足水分，分装，灭菌，备用。使用该培养基对pH值要求不严格，可以不测定。
配方二 综合马铃薯培养基
20%马铃薯煮汁 1000毫升
磷酸二氢钾 3克 硫酸镁 1.5克
葡萄糖 20克 维生素 10毫克
琼脂 18克
先配制20%马铃薯煮汁，方法同上。在煮汁中加入上述各种组分，加热溶解后补足水分，调整pH值到6。分装，灭菌，备用。 该培养基用于培养和保存灵芝、平菇、香菇等食用菌菌种。
Formula of common microbiological medium
(1) when separating bacteria, adding concentration of 50U/m in the culture medium; (2) adding 0.05% twelve alkanes in the samples when separating actinomycetes; (3) when separating fungi and yeast, adding penicillin to the culture medium, and (4) separating Rhizopus and Mucor, because the mycelium of these microbes is tendril; it is generally used to separate the single order. The culture medium of microbes all contain inhibition of it; 1, bacterial culture medium; formula a beef paste agar medium; beef cream 0.3 grams, peptone 1 grams, sodium chloride 0.5 grams, water 1000 ml; in the burning of 1000 ml;
(1) when the bacteria were isolated, adding 50U/ml to nystatin in the medium could inhibit the growth of mold and yeast.
(2) when separating actinomycetes, adding 0.05% sodium alkyl sulfonate (SDS) in the sample can not only inhibit the growth of bacteria, but also activate the germination of actinomycetes spore. The addition of norfloxacin (5mg/L) + nystatin (50mg/L) + penicillin (0.8mg/L) can also effectively inhibit bacteria and fungi without affecting the growth of actinomycetes.
(3) when mold and yeast were isolated, penicillin, streptomycin and tetracycline 30U/ml were added to the medium, and the growth of bacteria and actinomycetes could be inhibited.
(4) when Rhizopus and Mucor are separated, the mycelium of these microbes is easy to spread to pieces, and it is difficult to get the purified colony. It is usually added 0.1% deoxycholate or sorbitol to prevent the spread of mycelium in the culture medium, so that the colony grows small and tight.
The culture medium used for isolation of single purpose microbes contains inhibitors that inhibit other microorganisms. These special inhibitors are more troublesome in small laboratories and are now available to the base medium as long as they are directly added to the basic culture medium. Ampicillin, for example, is mainly used for the isolation of monospora hydrophila.
1. Bacterial culture medium
Formula one beef paste agar medium
0.3 grams of beef paste, 1 grams of peptone, 0.5 grams of sodium chloride, 1.5 grams of agar.
1000 milliliters of water
Add 100 ml of water to the beaker. Add beef paste, peptone and sodium chloride. Use crayons to mark the beaker and put them on the fire. When the components in the beaker are dissolved, add agar, stirring continuously to avoid sticking to the bottom. After the agar was completely dissolved, the pH value was adjusted to 7.2 ~ 7.6 with 10% hydrochloric acid or 10% sodium hydroxide, which was installed in the test tubes, added cotton plug, and sterilized with high pressure steam for 30 minutes.
Formula two potato culture medium
Take fresh beef heart (remove fat and blood vessels) 250 grams, with a knife finely chopped into minced meat, add 500 ml distilled water and 5 grams peptone. Mark the beaker, boil it, and use it to simmer for 2 hours. After filtration, the dried meat was filtered and the pH value of the filtrate was adjusted to about 7.5. 10 milliliters of broth and a small amount of dried beef heart in each test tube were sterilized.
Formula four Rhizobium culture medium
Glucose 10 gram hydrogen phosphate, two potassium and 0.5 grams
Calcium carbonate 3 grams of Magnesium Sulfate 0.2 grams
Yeast 0.4 gram agar 20 grams
Water 1000 ml 1% crystal violet solution 1 ml
First boil the agar with water and dissolve it, then add other components separately, stir it to dissolve it, separate it, sterilize it and reserve it.
2. Actinomycetes culture medium
Formula starch agar medium (Kao's medium)
Soluble starch, 2 grams of potassium nitrate 0.1 grams
Two K of potassium hydrogen phosphate and 0.05 grams of sodium chloride 0.05 grams
Magnesium Sulfate 0.05 grams of ferrous sulphate 0.001 grams
Agar 2 grams of water 1000 ml
First put the starch in a beaker, then use 5 ml of water to paste into paste. Pour 95 ml of water into the beaker, stir it up and add other medicines to dissolve it. Mark the beaker outside, add the agar to the boil and stir until the agar is completely dissolved. Adjust the pH value to 7.2 to 7.4.
Formula two flour agar medium
Flour 60 grams agar 20 grams
1000 milliliters of water
Turn the flour into a paste, add water to 500 ml, and cook on a slow fire for 30 minutes. Take another 500 ml of water, add it to the agar, heat it to boil, dissolve it, mix up the two fluid, replenish the water, adjust the pH value to 7.4, pack, sterilize and reserve.
3. Fungal culture medium
Formula one pizza (Sabouraud 's) medium
Peptone 10 grams agar 20 grams
Maltose 40 grams of water 1000 ml
After adding peptone and agar to water, heat and stir continuously, after the agar is dissolved, add 40 grams of maltose (or glucose), stir, dissolve it, then pack, sterilize, and spare.
This culture is commonly used for the cultivation of many kinds of fungi.
Formula Two Potato sugar agar medium
Rinse the potatoes and remove the skin. Cut 200 grams into small pieces, add 1000 ml of water, and boil for half an hour to fill up the water. Add 10 gram agar into the filtrate, add 20 grams of sugar after boiling and dissolve (used for cultivation of mould with sucrose, used for culture yeast to add glucose), supplement water, pack, sterilize and spare.
The pH value of this medium is adjusted to 7.2 to 7.4. Sugar in the formula, such as glucose, can also be used to culture actinomycetes and Bacillus.
Culture medium of formula three yellow bean sprout juice
100 grams of yellow bean sprout 15 grams of agar
Glucose 20 grams of water 1000 ml
Wash the yellow bean sprouts and boil for 30 minutes with water. Use gauze to filter, add agar in the filtrate, dissolve it and heat it into sugar, stir it to dissolve it, make up the water to 1000 milliliters, separate it, sterilize it and reserve it.
The pH value of this medium is adjusted to 7.2 to 7.4, which can be used to cultivate bacteria and actinomycetes.
Formula four pea agar medium
80 grain agar 5 grams of pea
200 milliliters of water
Add 80 pieces of dried peas and add water. Boil for 1 hours. After gauze filtering, add agar in the filtrate, boil to dissolve, separate, sterilize and reserve.
4. The culture medium of edible fungus strain
Formula one potato - sucrose agar medium
20% potato juice 1000 ml
Sucrose 20 gram agar 18 grams
Wash the potatoes and peel them and cut them into small pieces. Weigh 200 grams of potatoes, add 1000 ml of water, and boil for 20 minutes. In the filter juice to fill up to 1000 ml of water, that is 20% potato juice. Add the agar and sucrose into the potato juice, boil it, dissolve it, make up the moisture, separate it, sterilize it and reserve it. The use of the medium does not require strict determination of pH value.
Formula two comprehensive potato culture medium
20% potato juice 1000 ml
Potassium dihydrogen phosphate 3 grams of Magnesium Sulfate 1.5 grams
Glucose 20 grams of vitamin 10 mg
Agar 18 grams
20% potato juice is prepared in the same way. The above components were added to the boiling juice. After heating and dissolving, the moisture content was compensated and the pH value was adjusted to 6. Separate, sterilize, spare. The medium was used to cultivate and preserve edible fungi of Ganoderma lucidum, Pleurotus ostreatus and letinous edodes.