培養基之製備.doc

生物學實驗結報 組別:3 姓名:李隆益、王彥勳 一、培養基之製備 了解培養微生物所需之各種培養基並學習配製方法。液體培養基(Broth)中加入洋菜作為固化劑，即成固體培養基。固體培養基若分裝於培養皿中，即為平板培養基(Plate)。用燒杯(Beaker)先裝少量的蒸餾水，再按照配方逐項將藥品秤好，逐項分別放入燒杯中攪拌溶解，全部藥品均秤好，溶解均勻後，加水到所需要的體積。平板培養基：按照上項方法將培養基配好後，分裝於錐形瓶(flask)中。再加適量洋菜(agar一般為1.5~1.8%)，輕輕搖動，使洋菜分散。利用高壓蒸氣滅菌。滅好菌後，等溫度下降到50~60℃ (若有抗生素等不耐熱之物質，則於此時以過濾法加入)，手握錐形瓶輕輕搖動，以使錐形瓶中所有內容物混合均勻，且不起泡，倒入無菌培養皿中，凝固(洋菜在45℃開始凝固)即可。瓊脂平板每片瓊脂平板約15~20 ml配置片瓊脂平板需約00 ml). 2. 取一ml 錐形瓶。用量筒量ml的蒸餾水加入錐形瓶中。秤取加入錐形瓶中。再加入0ml的蒸餾水，溶解均勻。秤取agar g (1.5%~1.8%) 加入錐形瓶中。 用雙層錫箔紙蓋住瓶口，在錫箔紙上標示培養基名稱、組別、姓名。貼上滅菌膠帶後，送入高壓滅菌鍋滅菌15~20分鐘。由高壓滅菌鍋取出後，輕搖錐形瓶，使內容物混合均勻且不起泡。桌面及雙手噴酒精消毒。取出無菌培養皿，於底蓋標示培養基名稱、日期、組別、姓名。待溫度降至50~60℃左右(手握錐形瓶不燙)。點燃酒精燈，打開錫箔紙，瓶口過火，於酒精燈火焰附近（無菌空氣罩區域）將培養基倒入培養皿中。待培養基凝固，倒放入37℃培養箱中培養。隔天觀察是否有污染。沒有污染的培養基，即可放入冰箱中備用。Induction of mutations and Gram Stain 目的: 1. To test a hypothesis that mutations can be induced by ultraviolet light 2. To interpret results from bacterial mutagenesis experiments 原理: Natural selection is the agent in nature that determines whether a mutation is good or bad. A detrimental mutation (allele) would be selected against; i .e., organisms with the mutation would not function as well in the environment and would leave fewer offspring. Over time, the frequency of genotypes carrying a detrimental mutation should decrease in the population. The opposite would be true for a beneficial one. State a hypothesis to test in your experiment. It should relate the length of exposure to UV light to the amount of mutation expected. In this part of the lab topic, you will experimentally induce mutations in a bacterium, Serratia marcescens. Bacteria are good organisms to use in mutation studies because they are haploid. If a mutation occurs, it is expressed because there is not a second allele present to mask it. Mutations affecting viability in Serratia. Mutations can be induced by several means. Chemicals, called mutagens can change an organisms DNA, causing changes in hereditary information. Ultraviolet radiation has similar effects and will be used in th