常用微生物培養(yǎng)基配方大全
發(fā)布時(shí)間：
2022-11-16
作者：
常用微生物培養(yǎng)基配方大全
（1）分離細(xì)菌時(shí)，在培養(yǎng)基中加入濃度為50U/m；（2）分離放線菌時(shí)，在樣品中加入0.05%十二烷；（3）分離霉菌和酵母菌時(shí)，在培養(yǎng)基中加入青霉素、；（4）分離根霉和毛霉時(shí)，由于這些微生物的菌絲易蔓；一般用于分離單一目的微生物的培養(yǎng)基中均含有抑制其；1、細(xì)菌培養(yǎng)基；配方一牛肉膏瓊脂培養(yǎng)基；牛肉膏0.3克，蛋白胨1.0克，氯化鈉0.5克，；水1000毫升；在燒
（1）分離細(xì)菌時(shí)，在培養(yǎng)基中加入濃度為50U/ml制霉菌素，可以抑制霉菌和酵母菌的生長。
（2）分離放線菌時(shí)，在樣品中加入0.05%十二烷基磺酸鈉（SDS）不僅可以抑制細(xì)菌的生長，還能激活放線菌孢子的萌發(fā)。加入氟哌酸（5mg/L）+制霉菌素（50mg/L）+青霉素（0.8mg/L）也可以有效地抑制細(xì)菌和真菌，而不影響放線菌的生長。
（3）分離霉菌和酵母菌時(shí)，在培養(yǎng)基中加入青霉素、鏈霉素和四環(huán)素各30U/ml，可以抑制細(xì)菌和放線菌生長。
（4）分離根霉和毛霉時(shí)，由于這些微生物的菌絲易蔓延成片，難以得到純化的菌落，通常在培養(yǎng)基中添加0.1%去氧膽酸鈉或山梨醇防止菌絲蔓延，使菌落長得小而緊密。
一般用于分離單一目的微生物的培養(yǎng)基中均含有抑制其他微生物的抑制劑，這些專用的抑制劑在小型實(shí)驗(yàn)室配制較麻煩，現(xiàn)已有成品供應(yīng)，只要直接加入基礎(chǔ)培養(yǎng)基即可。如氨芐西林，主要用于分離親水氣單孢菌。
1、細(xì)菌培養(yǎng)基
配方一 牛肉膏瓊脂培養(yǎng)基
牛肉膏0.3克 ，蛋白胨1.0克，氯化鈉 0.5克，瓊脂 1.5克，
水 1000毫升
在燒杯內(nèi)加水100毫升，放入牛肉膏、蛋白胨和氯化鈉，用蠟筆在燒杯外作上記號(hào)后，放在火上加熱。待燒杯內(nèi)各組分溶解后，加入瓊脂，不斷攪拌以免粘底。等瓊脂完全溶解后補(bǔ)足失水，用10%鹽酸或10%的氫氧化鈉調(diào)整pH值到7.2～7.6,分裝在各個(gè)試管里，加棉花塞，用高壓蒸汽滅菌30分鐘。
配方二 馬鈴薯培養(yǎng)基
取新鮮牛心（除去脂肪和血管）250克，用刀細(xì)細(xì)剁成肉末后，加入500毫升蒸餾水和5克蛋白胨。在燒杯上做好記號(hào)，煮沸，轉(zhuǎn)用文火燉2小時(shí)。過濾，濾出的肉末干燥處理，濾液pH值調(diào)到7.5左右。每支試管內(nèi)加入10毫升肉湯和少量碎末狀的干牛心，滅菌，備用。
配方四 根瘤菌培養(yǎng)基
葡萄糖 10克 磷酸氫二鉀 0.5克
碳酸鈣 3克 硫酸鎂 0.2克
酵母粉 0.4克 瓊脂 20克
水 1000毫升 1%結(jié)晶紫溶液 1毫升
先把瓊脂加水煮沸溶解，然后分別加入其他組分，攪拌使溶解后，分裝，滅菌，備用。
2、放線菌培養(yǎng)基
配方一 淀粉瓊脂培養(yǎng)基（高氏培養(yǎng)基）
可溶性淀粉 2克 硝酸鉀 0.1克
磷酸氫二鉀 0.05克 氯化鈉 0.05克
硫酸鎂 0.05克 硫酸亞鐵 0.001克
瓊脂 2克 水 1000毫升
先把淀粉放在燒杯里，用5毫升水調(diào)成糊狀后，倒入95毫升水，攪勻后加入其他藥品，使它溶解。在燒杯外做好記號(hào)，加熱到煮沸時(shí)加入瓊脂，不停攪拌，待瓊脂完全溶解后，補(bǔ)足失水。調(diào)整pH值到7.2～7.4，分裝后滅菌，備用。
配方二 面粉瓊脂培養(yǎng)基
面粉 60克 瓊脂 20克
水 1000毫升
把面粉用水調(diào)成糊狀，加水到500毫升，放在文火上煮30分鐘。另取500毫升水，放入瓊脂，加熱煮沸到溶解后，把兩液調(diào)勻，補(bǔ)充水分，調(diào)整pH值到7.4，分裝，滅菌，備用。
3、真菌培養(yǎng)基
配方一 薩市（Sabouraud’s）培養(yǎng)基
蛋白胨 10克 瓊脂 20克
麥芽糖 40克 水 1000毫升
先把蛋白胨、瓊脂加水后，加熱，不斷攪拌，待瓊脂溶解后，加入40克麥芽糖（或葡萄糖），攪拌，使它溶解，然后分裝，滅菌，備用。
本培養(yǎng)菌是培養(yǎng)許多種類真菌所常用的。
配方二 馬鈴薯糖瓊脂培養(yǎng)基
把馬鈴薯洗凈去皮，取200克切成小塊，加水1000毫升，煮沸半小時(shí)后，補(bǔ)足水分。在濾液中加入10克瓊脂，煮沸溶解后加糖20克（用于培養(yǎng)霉菌的加入蔗糖，用于培養(yǎng)酵母菌的加入葡萄糖），補(bǔ)足水分，分裝，滅菌，備用。
把這培養(yǎng)基的pH值調(diào)到7.2～7.4，配方中的糖，如用葡萄糖還可用來培養(yǎng)放線菌和芽孢桿菌。
配方三 黃豆芽汁培養(yǎng)基
黃豆芽 100克 瓊脂 15克
葡萄糖 20克 水 1000毫升
洗凈黃豆芽，加水煮沸30分鐘。用紗布過濾，濾液中加入瓊脂，加熱溶解后放入糖，攪拌使它溶解，補(bǔ)足水分到1000毫升，分裝，滅菌，備用。
把這培養(yǎng)基的pH值調(diào)到7.2～7.4，可用來培養(yǎng)細(xì)菌和放線菌。
配方四 豌豆瓊脂培養(yǎng)基
豌豆 80粒 瓊脂 5克
水 200毫升
取80粒干豌豆加水，煮沸1小時(shí)，用紗布過濾后，在濾液中加入瓊脂，煮沸到溶解，分裝，滅菌，備用。
4、食用菌菌種培養(yǎng)基
配方一 馬鈴薯—蔗糖--瓊脂培養(yǎng)基
20%馬鈴薯煮汁 1000毫升
蔗糖 20克 瓊脂 18克
把馬鈴薯洗凈去皮后，切成小塊。稱取馬鈴薯小塊200克，加水1000毫升，煮沸20分鐘后，過濾。在濾汁中補(bǔ)足水分到1000毫升，即成20%馬鈴薯煮汁。在馬鈴薯煮汁中加入瓊脂和蔗糖，煮沸，使它溶解后，補(bǔ)足水分，分裝，滅菌，備用。使用該培養(yǎng)基對(duì)pH值要求不嚴(yán)格，可以不測(cè)定。
配方二 綜合馬鈴薯培養(yǎng)基
20%馬鈴薯煮汁 1000毫升
磷酸二氫鉀 3克 硫酸鎂 1.5克
葡萄糖 20克 維生素 10毫克
瓊脂 18克
先配制20%馬鈴薯煮汁，方法同上。在煮汁中加入上述各種組分，加熱溶解后補(bǔ)足水分，調(diào)整pH值到6。分裝，滅菌，備用。 該培養(yǎng)基用于培養(yǎng)和保存靈芝、平菇、香菇等食用菌菌種。
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.