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牛肉膏蛋白胨瓊脂培養(yǎng)基配制步驟

時間:2011/11/8閱讀:1412
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牛肉膏蛋白胨瓊脂培養(yǎng)基這是一種應用十分廣泛的天然培養(yǎng)基,其中的牛肉膏為微生物提供碳源、磷酸鹽和維生素,蛋白胨主要提供氮源和維生素,而NaCl提供無機鹽。

配制具體步驟:
1.稱量:按培養(yǎng)基配方比例依次準確地稱取牛肉膏、蛋白胨、NaCl放入燒杯中。牛肉膏常用玻棒挑取,放在小燒杯或表面皿中稱量,用熱水溶化后倒入燒杯。也可放在稱量紙上,稱量后直接放入水中,這時如稍微加熱,牛肉膏便會與稱量紙分離,然后立即取出紙片。蛋白胨很易吸潮,在稱取時動作要迅速。另外,稱藥品時嚴防藥品混雜,一把牛角匙用于一種藥品,或稱取一種藥品后,洗凈、擦干,再稱取另一藥品,瓶蓋也不要蓋錯。
2.溶化:在上述燒杯中可先加入少于所需要的水量,用玻棒攪勻,然后,在石棉網(wǎng)上加熱使其溶解。待藥品*溶解后,補充水分到所需的總體積。如果配制固體培養(yǎng)基,將稱好的瓊脂放入已溶化的藥品中,再加熱溶化,在瓊脂溶化的過程中,需不斷攪拌,以防瓊脂糊底使燒杯破裂。zui后補足所失的水分。
3.調(diào)pH:在未調(diào)pH前,先用精密pH試紙測量培養(yǎng)基的原始pH值,如果pH偏酸,用滴管向培養(yǎng)基中逐滴加入1mol/L NaOH,邊加邊攪拌,并隨時用pH試紙測其pH值,直至pH達7.6。反之,則用1mol/L HCl進行調(diào)節(jié)。注意pH值不要調(diào)過頭,以避免回調(diào),否則,將會影響培養(yǎng)基內(nèi)各離子的濃度。   對于有些要求pH值較的微生物,其pH的調(diào)節(jié)可用酸度計進行(使用方法,可參考有關說明書)。
4.過濾:趁熱用濾紙或多層紗布過濾,以利結(jié)果的觀察。一般無特殊要求的情況下,這一步可以省去(本實驗無需過濾)。
5.分裝:按實驗要求,可將配制的培養(yǎng)基分裝入試管內(nèi)或三角燒瓶內(nèi)。
6.加塞:培養(yǎng)基分裝完畢后,在試管口或三角燒瓶口上塞上棉塞,以阻止外界微生物進入培養(yǎng)基內(nèi)而造成污染,并保證有良好的通氣性能(棉塞制作方法附本實驗后面)。
7.包扎:加塞后,將全部試管用麻繩捆扎好,再在棉塞外包一層牛皮紙,以防止滅菌時冷凝水潤濕棉塞,其外再用一道麻繩扎好。用記號筆注明培養(yǎng)基名稱、組別、日期。三角燒瓶加塞后,外包牛皮紙,用麻繩以活結(jié)形式扎好,使用時容易解開,同樣用記號筆注明培養(yǎng)基名稱、組別、日期。

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