| 菌株簡(jiǎn)介 |
1. 菌株保存在20%甘油的培養(yǎng)基中，接到菌株后，請(qǐng)?zhí)羧【涸?x YT-N平板（2xYT medium containing 100 μg/ml nalidixic acid）上劃線(xiàn)，37℃過(guò)夜培養(yǎng)后，再次挑取單個(gè)菌落在Minimal Medium 平板上劃線(xiàn)，37℃過(guò)夜培養(yǎng)后即可使用，以便使纖毛充分表達(dá)，利于噬菌體感染； 2. 在Minimal Medium 平板上生長(zhǎng)的細(xì)菌，不宜在冰箱中放置過(guò)久再使用； 3. Minimal Medium plate 配制方法： Prepare stocks of the following: 1 M MgCl2?6H2O: Dissolve 20.33 g in distilled water to a final volume of 100 ml and autoclave. 1 M CaCl2?2H2O: Dissolve 14.7 g in distilled water to a final volume of 100 ml and autoclave. 1 M thiamine hydrochloride: Dissolve 33.73 g in distilled water to a final volume of 100 ml and sterilize using a 0.22 μm filter. 20% glucose: Dissolve 20 g of D-(+)-glucose (anhydrous) in distilled water to a final volume of 100 ml and sterilize by filtration through a 0.2 μm filter. Do not autoclave. In a 500 ml bottle, dissolve 6 g of Na2HPO4 (dibasic), 3 g of KH2PO4(monobasic) and 1 g of NH4Cl in distilled water to a final volume of 500 ml. adjust the pH to 7.4 with NaOH. In a separate 1 liter bottle, add distilled water to 15 g of Bacto-agar to a final Volume of 500 ml. Autoclave both bottles simultaneously to sterilize. Cool both bottles to 50-60°C and combine. Add1 ml of 1 M MgCl2?6H2O, 1 ml of 1 M CaCl2?2H2O, 1 ml of 1 M thiamine hydrochloride and 5 ml of 20% glucose. Pour plates immediately. |
