常用培養(yǎng)基的制備，滅菌與消毒實(shí)驗(yàn)
發(fā)布時(shí)間：
2022-12-27
作者：
即用型干粉培養(yǎng)基
1，實(shí)驗(yàn)?zāi)康模?nbsp;
了解培養(yǎng)基的配制原理；掌握配制培養(yǎng)基的一般方法和步驟；了解常見(jiàn)滅菌，清毒基本原理及方法，掌握干熱天菌，高壓蒸汽滅菌及過(guò)濾除菌的操作方法。
2，實(shí)驗(yàn)原理：
培養(yǎng)基是人工按一定比例配制的供微生物生長(zhǎng)繁殖和合成代謝產(chǎn)物所需要的營(yíng)養(yǎng)物質(zhì)的混合物。培養(yǎng)基的原材料可分為碳源，氮源，無(wú)機(jī)鹽，生長(zhǎng)因素和水。根據(jù)微生物的種類(lèi)和實(shí)驗(yàn)?zāi)康牟煌囵B(yǎng)基也有不同的種類(lèi)和配制方法。
牛肉膏蛋白胨培養(yǎng)基是一種應(yīng)用最廣泛和最普通的細(xì)菌基礎(chǔ)培養(yǎng)基，有時(shí)又稱為普通培養(yǎng)基。由于這種培養(yǎng)基中含有一般細(xì)胞生長(zhǎng)繁殖所需要的最基本的營(yíng)養(yǎng)物質(zhì)，所以可供微生物生長(zhǎng)繁殖之用。
干熱天菌，高壓蒸汽滅菌方法主要是通過(guò)升溫使蛋白質(zhì)變性從而達(dá)到殺死微生物的效果。
3，試劑與器材：
1.器材 試管，三角瓶，燒杯，量筒，玻璃棒，培養(yǎng)基，分裝器，天平，牛角匙，高壓蒸汽滅菌鍋，pH度紙，棉花，牛皮紙，記號(hào)筆，麻繩，紗布，吸管，培養(yǎng)皿，電烘箱，注射器，微孔濾膜過(guò)濾器，鑷子等。
2.試劑 牛肉膏，蛋白胨，NaCl，瓊脂。
4，實(shí)驗(yàn)內(nèi)容：
1.稱量—溶化—調(diào)pH—過(guò)濾—分裝—加塞—包扎—滅菌—無(wú)菌檢查。
2.干熱滅菌：裝入待滅菌物品—升溫—恒溫—降溫—開(kāi)箱取物。 
3.高壓蒸汽滅菌：加水—裝物品—加蓋—加熱—排冷空氣—加壓—恒壓—降壓回零—排汽—取物—無(wú)菌檢查。
4.過(guò)濾除菌：組裝滅菌—連接—壓濾—無(wú)菌檢查—清洗滅菌。
5，關(guān)鍵步驟及注意事項(xiàng)：
1.要嚴(yán)格按配方配制。
2.調(diào)pH不要過(guò)頭。
3.干熱滅菌要注意物品不要堆放過(guò)緊，注意溫度的時(shí)間控制，70ºC以下放物，取物。
4.高壓滅菌要注意物品不要過(guò)多，加熱后排除冷空氣，到時(shí)降壓回零取物。
5.過(guò)濾除菌要注意各部件滅菌，壓濾時(shí)，壓力要適當(dāng)，不可太猛太快，濾膜要注意清洗保存。
Preparation, sterilization and disinfection of commonly used medium
1, the purpose of the experiment is:
Understand the preparation principle of the medium, master the general methods and steps of preparing the medium, understand the basic principles and methods of common sterilization and detoxification, master the operating methods of dry and hot days, high-pressure steam sterilization and filtration sterilization.
2, the experimental principle:
Medium is a mixture of nutrients for microbial growth, reproduction and metabolites. The raw materials of the culture medium can be divided into carbon source, nitrogen source, inorganic salt, growth factor and water. According to the different types of microorganisms and experimental purposes, there are different kinds of media and preparation methods.
Beef extract peptone culture medium is one of the most widely used and common bacterial base medium, sometimes known as ordinary medium. Because this kind of medium contains the most basic nutrients for the growth and reproduction of ordinary cells, it can be used for the growth and reproduction of microorganisms.
Dry-hot days bacteria, high-pressure steam sterilization method is mainly through heating protein denaturation to achieve the effect of killing microorganisms.
3, reagents and equipment:
1. Equipment test tube, triangle bottle, beaker, measuring barrel, glass rod, culture medium, sub-packer, balance, horn spoon, high-pressure steam sterilization pot, pH paper, cotton, kraft paper, marker pen, hemp rope, gauze, straw, culture dish, electric oven, syringe, microporous filter, tweezers, etc.
2. reagent beef paste, peptone, NaCl, agar.
4, experimental contents:
1. weighing - dissolving - adjusting pH - Filtration - packing - plus - dressing - sterilization - sterility test.
2. dry heat sterilization: loading the product to be sterilized - warming up - constant temperature - cooling - opening the box.
3. Sterilization by high pressure steam: adding water, loading, covering, heating, discharging cold air, pressurizing, constant pressure, depressurizing back to zero, discharging steam, extracting, sterilization inspection.
4. filtration sterilization: assembly sterilization - connection - pressure filtration - sterility test - cleaning and sterilization.
5, the key steps and matters needing attention:
1., we must strictly prepare the formula.
2. don't overdo pH.
3. Dry-heat sterilization should pay attention to the items do not stack too tight, pay attention to the time control of temperature, 70 C below the place, take things.
4. High-pressure sterilization should pay attention to not too many articles, after heating, cold air is removed, when the time comes to reduce the pressure back to zero.
5. Pay attention to the sterilization of each component in filtration, pressure should be appropriate, not too violent and too fast, filter membrane should pay attention to cleaning and preservation.